over time using live cell microscopy. The time post infection is indicated in the lower left corner.
|
|
|
- Emily Lamb
- 6 years ago
- Views:
Transcription
1 Title of file for HTML: Supplementary Information Description: Supplementary Figures and Supplementary Table Title of file for HTML: Supplementary Movie 1 Description: Fusion of NBs. BSR cells were infected by rcvsn2c-p-mcherry and imaged over time using live cell microscopy. The time post infection is indicated in the lower left corner. Title of file for HTML: Supplementary Movie 2 Description: Fusion of NBs. BSR cells were infected by rcvsn2c-p-mcherry and imaged over time using live cell microscopy. The time post infection is indicated in the lower left corner. Title of file for HTML: Supplementary Movie 3 Description: A spherical cytoplasmic bubble crosses a NB. BSR cells were infected by rcvsn2c-p-mcherry and imaged over time using live cell microscopy. The time post infection is indicated in the lower left corner. Title of file for HTML: Supplementary Movie 4 Description: NBs are sensitive to a hypotonic chock. BSR cells were infected by rcvsn2c-pmcherry and imaged over time using live cell microscopy. A hypotonic shock was applied 5 minutes after the beginning of the movie (~18h p.i.). The time post-infection is displayed in the lower left corner. Title of file for HTML: Supplementary Movie 5 Description: NBs and SGs are non-miscible liquid organelles. U373-MG cells were transiently transfected with pg3bp-egfp (to visualize SGs in white). 1h post transfection, they were infected with the recombinant virus rcvsn2c-p-mcherry (to visualize NBs in red). At 16h p.i., both NBs and SGs were imaged. The time post-infection is displayed in the lower left corner. Title of file for HTML: Supplementary Movie 6 Description: RNPs are ejected from NBs. BSR cells were infected by rcvsn2c-p-mcherry BSR cells were infected by rcvsn2c-p-mcherry. At 16h p.i., NBs and RNPs were imaged. The time post-infection is displayed in the lower left corner. Title of file for HTML: Supplementary Movie 7 Description: RNPs are ejected from NBs. BSR cells were infected by rcvsn2c-p-mcherry BSR cells were infected by rcvsn2c-p-mcherry. At 16h p.i., NBs and RNPs were imaged. The time of acquisition is displayed in the lower left corner. Title of file for HTML: Supplementary Movie 8 Description: RNPs are mobile in the cytosol of infected cells. BSR cells were infected by rcvsn2c-p-mcherry. At 20h p.i., NBs and RNPs were imaged. The time post-infection is displayed in the lower left corner. Title of file for HTML: Supplementary Movie 9
2 Description: Nocodazole inhibits RNPs movement. Nocodazole (2 µm) was added 1h before and kept all along infection of BSR cells by rcvsn2c-p-mcherry. At ~23h p.i., NBs and RNPs were imaged. The time post-infection is displayed in the lower left corner. Title of file for HTML: Supplementary Movie 10 Description: RNPs are transported along microtubules. BSR cells were co-infected with rcvs N2C-P-mCherry and a modified baculovirus encoding human tubulin-gfp. Cells were imaged after 16h p.i. Title of file for HTML: Supplementary Movie 11 Description: No ejection is observed from N-P like structures in the minimal system. BSR- T7/5 cells were co-transfected with plasmids ptit-p-mcherry and ptit-n. Cells were imaged at ~24h post-transfection. Title of file for HTML: Supplementary Movie 12 Description: No ejection is observed from N-P like structures in the minimal system (when cotranfected with ptit M). BSR-T7/5 cells were co-transfected with plasmids ptit-p-mcherry, ptit-n and ptit-m. Cells were imaged at ~24h post-transfection. Title of file for HTML: Peer Review File Description:
3 Supplementary Information Supplementary Figure 1 (related to Figure 2): rcvsn2c-p-mcherry behaves as wild type CVS. BSR cells were infected with rcvsn2c-p-mcherry at a MOI of 0.5 and fixed at 8 and 24h p.i. (A) Confocal analysis was performed after staining with a mouse monoclonal anti-n antibody followed by incubation with Alexa-488 donkey anti-mouse IgG. DAPI was used to stain the nuclei. (B) Confocal analysis revealing the basal localization of small dots and the median localization of inclusions in RABV infected cells at 24h p.i. The analysis was performed after staining as in (A). Scale bars correspond to 15 µm.
4 Normalized Fluo C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C Time (sec.) Supplementary Figure 2 (related to Figure 2E): Fluorescence recovery after photobleaching (FRAP) of P-mCherry expressed after transfection in BSR cells Cytosolic P-mCherry was photobleached 24h after transfection of ptit-p-mcherry plasmid. The FRAP curves corresponding to each of the 11 FRAP events are shown. They have been used to obtain the mean curve of figure 2E.
5 Normalized Fluo C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C Time (sec.) Supplementary Figure 3 (related to Figure 2F): Fluorescence recovery after photobleaching (FRAP) of cytosolic P-mCherry expressed in BSR cells infected by rcvsn2cδg-p-mcherry. Cytosolic P-mCherry was photobleached 16h p.i. The FRAP curves corresponding to each of the 13 FRAP events are shown. They have been used to obtain the mean curve of figure 2F.
6 Normalized Fluo C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C Time (sec.) Supplementary Figure 4 (related to Figure 2G): Fluorescence recovery after photobleaching (FRAP) of P-mCherry in NBs formed in BSR cells infected by rcvsn2cδg-p-mcherry. Cytosolic P-mCherry located in NBs was photobleached 16h p.i. The FRAP curves corresponding to each of the 12 FRAP events are shown. They have been used to obtain the mean curve of figure 2G.
7 Supplementary Figure 5 (related to Figure 5): Images of BSR cells co-infected with rcvs N2C-P-mCherry and a modified baculovirus encoding human tubulin-gfp (Celllight Tubulin-GFP) before (left) and after (right) deconvolution using the Huygens Imaging software.
8 Supplementary Figure 6 (related to Figure 6A): Dependence of NB-like structures formation on the stoechiometry of ptit-n and ptit-p plasmids. BSR-T7/5 cells were co-transfected with different ratios of ptit-p and ptit-n (keeping constant the total amount of DNA to 800ng for cells). N was revealed with a mouse monoclonal anti-n antibody followed by incubation with Alexa-488 donkey antimouse IgG and P was revealed with a rabbit polyclonal anti-p antibody followed by incubation with Alexa-568 donkey antirabbit IgG. DAPI was used to stain the nuclei. Scale bars correspond to 15 µm.
9 Normalized Fluo C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 C15 C16 C17 C18 C19 C20 C Time (sec.) Supplementary Figure 7 (related to Figure 6B): Fluorescence recovery after photobleaching (FRAP) of P-mCherry in NB-like inclusions formed in BSR-T7/5 cells co-transfected by ptit-p-mcherry and ptit-n. Cytosolic P-mCherry located in NB-like inclusions was photobleached 24h post-transfection. The FRAP curves corresponding to each of the 21 FRAP events are shown. They have been used to obtain the mean curve of figure 6B.
10 Supplementary Figure 8 (related to Figure 6) (A) Localization of Hsp70 in NBs and N-P inclusions in the minimal system BSR T7/5 cells were infected with CVS strain (RABV) at a MOI of 1 and fixed at 16h p.i (top row) or co-transfected with plasmids ptit-p and ptit-n (in equimolar concentration) and fixed at 24h post-transfection (bottom row). Confocal analysis was performed after staining with a rabbit polyclonal anti-p antibody (P) followed by incubation with Alexa-488 donkey, and a mouse MAb anti-hsp70 antibody (Hsp70) followed by incubation with Alexa 568 donkey anti-mouse IgG. (B) Localization of FAK-GFP in NBs and NB-like structures (N-P inclusions) BSR T7/5 cells were transfected with plasmid encoding FAK-GFP, then infected at an MOI of 1 for 24 h (top row) or co-transfected with plasmids ptit-p and ptit-n and plasmid encoding FAK-GFP (in equimolar concentrations) (Bottom row). Cells were analyzed by confocal microscopy after staining with a mouse monoclonal anti-n antibody followed by incubation with Alexa 568 anti-mouse IgG. The scale bars correspond to 15 µm. DAPI was used to stain the nuclei.
11 Supplementary Figure 9 (related to Figure 6 D): Role of P phosphorylation in NB-like structures formation BSR-T7/5 cells were co-transfected for 24h, with plasmids ptit-n and the indicated ptit-p mutants (in equimolar concentration). N was revealed with a mouse monoclonal anti-n antibody followed by incubation with Alexa-488 donkey anti-mouse IgG and P was revealed with a rabbit polyclonal anti-p antibody followed by incubation with Alexa-568 donkey antirabbit IgG. DAPI was used to stain the nuclei. Scale bars correspond to 15 µm.
12 * PNTD P-N 0 * * * * * CVS 1 MSKIFVNPSAIRAGLADLEMAEETVDLINRNIEDNQAHLQGEPIEVDNLPEDMKRLHLDD LBV 1 MSKGLIHPSAIRSGLVDLEMAEETVDLVHKNLADSQAHLQGEPLNVDSLPEDMRKMRLTN MOK 1 MSKDLVHPSLIRAGIVELEMAEETTDLINRTIESNQAHLQGEPLYVDSLPEDMSRLRIED DUV 1 MSKIFINPSDIRSGLADLEMAEETVELVNRNMEDSQAHLQGVPIDVETLPEDIQRLHITD EBL1 1 MSKIFVNPSALRSGLADLEMAEETVDLVNKNMEDSQAHLQGIPIDVETLPEDIKRLRIAD EBL2 1 MSKIFVNPSAIRAGLADLEMAEETVDLVNKNIEDNQAHLQGEPIEVDALPEDMSKLQISE ABL 1 MSKIFVNPSAIRAGMADLEMAEETVDLINRNIEDNQAHLQGEPIEVDSLPEDIKKLDISE IDD1 DD * * * * * * CVS 61 EKSSNLGEMVRVGEGKYREDFQMDEGEDPNLLFQSYLDNVGVQIVRQMRSGERFLKIWSQ LBV 61 APSEREIIEEDEEEYSSEDEYYLSQGQDPMVPFQNFLDELGTQIVRRMKSGDGFFKIWSA MOK 61 KSRRTKTEEEERDEGSSEEDNYLSEGQDPLIPFQNFLDEIGARAVKRLKTGEGFFRVWSA DUV 61 PQASLRQDMVDEQKHQEDEDFYLTGRENPLSPFQTHLDAIGLRIVRKMKTGEGFFKIWSQ EBL1 61 YKQGQREEDASRQEEGEDEDFYMTESENSYVPLQSYLDAVGMQIVRKMKTGDGFFKIWAQ EBL2 61 RRPAQFTDNTGGKEEGSDEDFYMAESEDPYIPLQSYLEGVGIQLVRQMKTGERFFKIWSQ ABL 61 GRSKSSADNPQDVDCRMSEDFQMDEVEDPNIQFQSYLDNIGIQIVRKMRTGERFFKIWSQ IDD2 * * * * * * CVS 121 TVEEIVSYVTVNFPNPPRRSSEDKSTQTTGRELK-KETTSAFSQRESQPSKARMVAQVAP LBV 121 ASEDIKGYVLSTFMKPETQATVSKPTQTDSLSVPRPSQGYTSVPRDKPSNSESQGGGVKP MOK 121 LSDDIKGYVSTNIMTSGERDTKSIQIQTEPTASVSSGNESRHDSESMHDPNDKKDHTPDH DUV 121 AVEDIVSYVALNFSIPVNKLFEDKSTQTVTEKSQQASASSAPNRHEKSSQNARVNSKDAS EBL1 121 AVEDIVSYVATNFPAPVNKLQADKSTQTTLEKVKQAVSSSAPNKREGPSSNMNLDSQESS EBL2 121 AVEEIISYVTVHFPMPLGKSTEDKSTQTPEEKFK-PSPQQAVTKKESQSSKIKTISQESS ABL 121 TVEEIISYVGVNFPNQSGKTTENKSTQTTPKKVK-TEPSSTPAKRSDQLSKTEMAAKTAS PCTD P-N ARN * * * * * * CVS 180 GPPALEWSATNEEDDLSVEAEIAHQIAESFSKKYKFPSRSSGIFLYNFEQLKMNLDDIVK LBV 181 KKVQKSEWTRDTDEISDIEGEVAHQVAESFSKKYKFPSRSSGIFLWNFEQLKMNLDDIVK MOK 181 DVVPDIESSTDKGEIRDIEGEVAHQVAESFSKKYKFPSRSSGIFLWNFEQLKMNLDDIVK DUV 181 GPAALDWTASNEADDESVEAEIAHQIAESFSKKYKFPSRSSGIFLWNFEQLKMNLDEIVR EBL1 181 GPPGLDWAASNDEDDGSIEAEIAHQIAESFSKKYKFPSRSSGIFLWNFEQLKMNLDDIVR EBL2 180 GPPALEWSTTNDEENASVEAEIAHQIAESFSKKYKFPSRSSGIFLFNFEQLKMNLDDIVK ABL 180 GPPALEWPTTNDEDDVSVEAEIAHQIAESFSKKYKFPSRSSGIFLYNFEQLKMNLDDIVK * * * * * * CVS 240 EAKNVPGVTRLAHDGSKIPLRCVLGWVALANSKKFQLLVEADKLSKIMQDDLNRYTSC LBV 241 TSMNVPGVDKIAEKGGKLPLRCILGFVSLDSSKRFRLLADTDKVARLMQDDIHNYMTRIE MOK 241 AAMNVPGVERIAEKGGKLPLRCILGFVALDSSKRFRLLADNDKVARLIQEDINSYMARLE DUV 241 EVKEIPGVIKMAKDGMKLPLRCMLGGVASTHSRRFQILVNPEKLGKVMQEDLDKYLTY EBL1 241 EVKGIPGVTRMARDGMKLPLRCMLGSVASNHSKRFQILVNSAKLGKLMQDDLNRYLAY EBL2 240 EAKKIPGVVRLAQDGFRLPLRCILGGVGSVNSKKFQLLVNSDKLGKIMQDDLNRYLAY ABL 240 EAKSVPGVTSLARDGLRLPLRCILGWVGSSHSKKFQLLVGSEKLNKIMQDDLNRYMSC LBV 301 EIDHN MOK 301 EAE Supplementary Figure 10 (related to Figure 6C, 6D, 6E and the discussion). Sequence alignment of P proteins of viruses, representative of the 7 genotypes of lyssaviruses. CVS (Rabies virus strain, genotype 1, Swiss-prot: P22363), LBV (Lagos Bat virus, genotype 2,
13 Swiss-Prot: O56773), MOK (Mokola Virus, genotype 3, Swiss-Prot: P0C569), DUV (Duvenhage virus, genotype 4, Swiss-Prot: O56774), EBL1 (European Bat Lyssavirus 1, genotype 5, GenBank: AAC04587), EBL2 (European Bat Lyssavirus 2, genotype 6, Swiss- Prot: A4UHQ4), ABL (Australian Bat Lyssa virus, genotype 7, Swiss-Prot: Q91RE5). Conserved residues are in cyan boxes whereas similar ones are in yellow (overall aa sequence conservation is 32 %). The black dashes show gaps. Rectangular boxes delimit protein domains. PNTD P-N 0 : N-terminal domain which binds to N 0 ; IDD1 and IDD2: intrinsically disordered domains; DD: dimerization domain; PCTD P-N ARN : C-terminal domain which binds to RNA-associated N protein. CVS residues 132 to 150 which are required for the phase transition are in bold. Cytosolic P-mCherry in transfected cells (Fig. 2E) Cytosolic P-mCherry in infected cells (Fig. 2F) P-mCherry in NBs in infected cells (Fig.2G) P-mCherry in inclusions in cells co-expressing N and P (Fig.6B) A fast k fast -1 (sec ) t 1/2 = ln2/k fast (sec) Diffusion coefficient associated with the fast phase (µm²/s) A slow k slow -1 (sec ) 0,5605 1,6903 0,410 ~5 0,43 0, ,5933 1,7401 0,4 ~5 0,3945 0,1504 0,004 0,4189 0,1343 5,2 ~0,4 0,4985 0,0109 0,0066 0,1477 0,123 5,6 ~0,36 0,5691 0, y 0 Supplementary Table 1 (related to Figure 2E, 2G, 2F and 6B) : FRAP parameters obtained by modeling the curves with a double exponential equation. The FRAP curves were fitted to a double exponential equation: ( ) ( ) ( ) The fitted values of the parameters, the deduced characteristic recovery time of the fast phase and a corresponding diffusion coefficient are indicated.
T H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Kanaani et al., http://www.jcb.org/cgi/content/full/jcb.200912101/dc1 Figure S1. The K2 rabbit polyclonal antibody is specific for GAD67,
Supplemental figures Supplemental Figure 1: Fluorescence recovery for FRAP experiments depicted in Figure 1.
Supplemental figures Supplemental Figure 1: Fluorescence recovery for FRAP experiments depicted in Figure 1. Percent of original fluorescence was plotted as a function of time following photobleaching
Supplemental Information. A Versatile Tool for Live-Cell Imaging. and Super-Resolution Nanoscopy Studies. of HIV-1 Env Distribution and Mobility
Cell Chemical Biology, Volume 24 Supplemental Information A Versatile Tool for Live-Cell Imaging and Super-Resolution Nanoscopy Studies of HIV-1 Env Distribution and Mobility Volkan Sakin, Janina Hanne,
Supplementary Figure 1. α-synuclein is truncated in PD and LBD brains. Nature Structural & Molecular Biology: doi: /nsmb.
Supplementary Figure 1 α-synuclein is truncated in PD and LBD brains. (a) Specificity of anti-n103 antibody. Anti-N103 antibody was coated on an ELISA plate and different concentrations of full-length
T H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Rainero et al., http://www.jcb.org/cgi/content/full/jcb.201109112/dc1 Figure S1. The expression of DGK- is reduced upon transfection
T H E J O U R N A L O F C E L L B I O L O G Y
Supplemental material Thompson et al., http://www.jcb.org/cgi/content/full/jcb.200909067/dc1 T H E J O U R N A L O F C E L L B I O L O G Y Figure S1. Modification-specific antibodies do not detect unmodified
SUPPLEMENTARY INFORMATION
doi: 10.1038/nature06147 SUPPLEMENTARY INFORMATION Figure S1 The genomic and domain structure of Dscam. The Dscam gene comprises 24 exons, encoding a signal peptide (SP), 10 IgSF domains, 6 fibronectin
Supplementary Figure Legends
Supplementary Figure Legends Supplementary Fig. 1. The third PDZ domain of PAR-3 and the C-terminal PDZ domain binding motif of VE-cadherin mediate the recruitment of PAR-3 to cell-cell contacts in cells.
Supplementary Figure S1
Supplementary Figure S1 Figure S1. CENP- FP constructs target to centromeres irrespective of site of tagging. FP- CENP and CENP- FP constructs were transfected into U2OS cells and counterstained with anti-
SUPPLEMENTARY INFORMATION
Nanomechanical mapping of first binding steps of a virus to animal cells David Alsteens, Richard Newton, Rajib Schubert, David Martinez-Martin, Martin Delguste, Botond Roska and Daniel J. Müller This PDF
SUPPLEMENTARY INFORMATION FIGURE LEGENDS
SUPPLEMENTARY INFORMATION FIGURE LEGENDS Fig. S1. Radiation-induced phosphorylation of Rad50 at a specific site. A. Rad50 is an in vitro substrate for ATM. A series of Rad50-GSTs covering the entire molecule
Table S1. List of DNA constructs and primers, part 1 Construct
SUPPLEMENTARY TABLES: Table S1. List of DNA constructs and primers, part 1 Construct Comment (Expressed protein) Vector, PCR primers and template or source of Restriction Sites name Resistance construct
Supplementary Figure Legends
Supplementary Figure Legends Figure S1 gene targeting strategy for disruption of chicken gene, related to Figure 1 (f)-(i). (a) The locus and the targeting constructs showing HpaI restriction sites. The
supplementary information
DOI: 10.1038/ncb2172 Figure S1 p53 regulates cellular NADPH and lipid levels via inhibition of G6PD. (a) U2OS cells stably expressing p53 shrna or a control shrna were transfected with control sirna or
Figure legends for supplement
Figure legends for supplement Supplemental Figure 1 Characterization of purified and recombinant proteins Relevant fractions related the final stage of the purification protocol(bingham et al., 1998; Toba
Figure 1: TDP-43 is subject to lysine acetylation within the RNA-binding domain a) QBI-293 cells were transfected with TDP-43 in the presence or
Figure 1: TDP-43 is subject to lysine acetylation within the RNA-binding domain a) QBI-293 cells were transfected with TDP-43 in the presence or absence of the acetyltransferase CBP and acetylated TDP-43
Supplementary Figure S1
Supplementary Figure S1 Supplementary Figure S1 Subcellular localization of Rab6a and LIS1 in the DRG neurons. (a) Exogenous expression of EGFP-Rab6a(wild-type) in the DRG neurons by the Neon transfection
T H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Nakajima and Tanoue, http://www.jcb.org/cgi/content/full/jcb.201104118/dc1 Figure S1. DLD-1 cells exhibit the characteristic morphology
Firefly luciferase mutants as sensors of proteome stress
Nature Methods Firefly luciferase mutants as sensors of proteome stress Rajat Gupta, Prasad Kasturi, Andreas Bracher, Christian Loew, Min Zheng, Adriana Villella, Dan Garza, F Ulrich Hartl & Swasti Raychaudhuri
Supplemental Data. Farmer et al. (2010) Plant Cell /tpc
Supplemental Figure 1. Amino acid sequence comparison of RAD23 proteins. Identical and similar residues are shown in the black and gray boxes, respectively. Dots denote gaps. The sequence of plant Ub is
Supplementary Table 1. The Q-PCR primer sequence is summarized in the following table.
Supplementary Table 1. The Q-PCR primer sequence is summarized in the following table. Name Sequence (5-3 ) Application Flag-u ggactacaaggacgacgatgac Shared upstream primer for all the amplifications of
(phosphatase tensin) domain is shown in dark gray, the FH1 domain in black, and the
Supplemental Figure 1. Predicted Domain Organization of the AFH14 Protein. (A) Schematic representation of the predicted domain organization of AFH14. The PTEN (phosphatase tensin) domain is shown in dark
SUPPLEMENTARY INFORMATION
DOI: 10.1038/ncb3363 Supplementary Figure 1 Several WNTs bind to the extracellular domains of PKD1. (a) HEK293T cells were co-transfected with indicated plasmids. Flag-tagged proteins were immunoprecipiated
Supplementary Information
Supplementary Information Supplementary Figure 1: Over-expression of CD300f in NIH3T3 cells enhances their capacity to phagocytize AC. (a) NIH3T3 cells were stably transduced by EV, CD300f WT or CD300f
Supplementary Materials for
www.sciencesignaling.org/cgi/content/full/9/429/ra54/dc1 Supplementary Materials for Dephosphorylation of the adaptor LAT and phospholipase C by SHP-1 inhibits natural killer cell cytotoxicity Omri Matalon,
Supplementary Figure 1. Confirmation of sirna in PC3 and H1299 cells PC3 (a) and H1299 (b) cells were transfected with sirna oligonucleotides
Supplementary Figure 1. Confirmation of sirna in PC3 and H1299 cells PC3 (a) and H1299 (b) cells were transfected with sirna oligonucleotides targeting RCP (SMARTPool (RCP) or two individual oligos (RCP#1
Supporting Information
Supporting Information Stavru et al. 0.073/pnas.357840 SI Materials and Methods Immunofluorescence. For immunofluorescence, cells were fixed for 0 min in 4% (wt/vol) paraformaldehyde (Electron Microscopy
Nature Biotechnology: doi: /nbt Supplementary Figure 1
Supplementary Figure 1 High dynamic range (HDR) imaging of live cells with spinning disk confocal microscopy. During image acquisition, z-sections, encompassing most of the cellular volume, are captured
Recruitment of Grb2 to surface IgG and IgE provides antigen receptor-intrinsic costimulation to class-switched B cells
SUPPLEMENTARY FIGURES Recruitment of Grb2 to surface IgG and IgE provides antigen receptor-intrinsic costimulation to class-switched B cells Niklas Engels, Lars Morten König, Christina Heemann, Johannes
Supplementary Figure 1 Structural modeling and purification of V. cholerae ABH. (a) The migration of the purified rabh and catalytically inactive
Supplementary Figure 1 Structural modeling and purification of V. cholerae ABH. (a) The migration of the purified rabh and catalytically inactive variants rabhs, rabhd, and rabhh are shown on 12% SDS-PAGE
JCB. Supplemental material THE JOURNAL OF CELL BIOLOGY. Prospéri et al.,
Supplemental material JCB Prospéri et al., http://www.jcb.org/cgi/content/full/jcb.201501018/dc1 THE JOURNAL OF CELL BIOLOGY Figure S1. Myo1b Tail interacts with YFP-EphB2 coated beads and genistein inhibits
Supplemental Information. Pacer Mediates the Function of Class III PI3K. and HOPS Complexes in Autophagosome. Maturation by Engaging Stx17
Molecular Cell, Volume 65 Supplemental Information Pacer Mediates the Function of Class III PI3K and HOPS Complexes in Autophagosome Maturation by Engaging Stx17 Xiawei Cheng, Xiuling Ma, Xianming Ding,
GFP CCD2 GFP IP:GFP
D1 D2 1 75 95 148 178 492 GFP CCD1 CCD2 CCD2 GFP D1 D2 GFP D1 D2 Beclin 1 IB:GFP IP:GFP Supplementary Figure 1: Mapping domains required for binding to HEK293T cells are transfected with EGFP-tagged mutant
Supplementary Figures
Supplementary Figures Supplementary Figure S1. Inhibition kinetics of p53/hdm2 binding induced by Nutlin 3 on live cells. Reproducibility of the p53/hdm2 binding disruption kinetics induced by Nutlin 3
Supplementary Figure 1. Espn-1 knockout characterization. (a) The predicted recombinant Espn-1 -/- allele was detected by PCR of the left (5 )
Supplementary Figure 1. Espn-1 knockout characterization. (a) The predicted recombinant Espn-1 -/- allele was detected by PCR of the left (5 ) homologous recombination arm (left) and of the right (3 )
indicated numbers of pups at day of life (DOL) 10, or embryonic day (ED) B. Male mice of
SUPPLEMENTRY FIGURE LEGENDS Figure S1. USP44 loss leads to chromosome missegregation.. Genotypes obtained from the indicated numbers of pups at day of life (DOL) 10, or embryonic day (ED) 13.5.. Male mice
Supplementary Information
Supplementary Information Figure S1. Transiently overexpressed ECFP-TIAF1/EYFP-TIAF1 causes apoptosis of Mv1Lu cells. Mv1Lu cells were transfected by electroporation with ECFP, EYFP, ECFP-TIAF1, EYFP-
Supplementary Information. A novel human endogenous retroviral protein inhibits cell-cell fusion. Supplementary Figures:
Supplementary Information A novel human endogenous retroviral protein inhibits cell-cell fusion Jun Sugimoto, Makiko Sugimoto, Helene Bernstein, Yoshihiro Jinno and Danny J. Schust Supplementary Figures:
SUPPLEMENTARY INFORMATION
SUPPLEMENTARY INFORMATION 1 Supplementary Figure S1. Proportion of HEK293T transfectants showing fluorescent foci after heat sho Blinded slides were counted for the appearance of fluorescent foci of tagrfp-t
SUPPLEMENTARY INFORMATION
doi: 10.1038/nature06721 SUPPLEMENTARY INFORMATION. Supplemental Figure Legends Supplemental Figure 1 The distribution of hatx-1[82q] in Cos7 cells. Cos7 cells are co-transfected with hatx-1[82q]-gfp (green)
SUPPLEMENTARY INFORMATION
DOI: 1.138/ncb37 a Supplementary Figure 1 Dapi EU γh2ax Overlay b Dapi NBS1-GFP Firbillarin Overlay c Pre-laser 2 min 5 min 1 min 2 min 3 min 4 min 5 min 6 min 7 min d Dapi GFP-MRE11 γh2ax Overlay Supplementary
SUPPLEMENTARY INFORMATION
DOI:.38/ncb327 a b Sequence coverage (%) 4 3 2 IP: -GFP isoform IP: GFP IP: -GFP IP: GFP Sequence coverage (%) 4 3 2 IP: -GFP IP: GFP 33 52 58 isoform 2 33 49 47 IP: Control IP: Peptide Sequence Start
T H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Craft et al., http://www.jcb.org/cgi/content/full/jcb.201409036/dc1 Figure S1. GFP -tubulin interacts with endogenous -tubulin. Ciliary
Nature Immunology: doi: /ni Supplementary Figure 1
Supplementary Figure 1 Sox2 localizes in neutrophils of mouse spleen. (a) Microscopy analysis of Sox2 and MPO in wild-type mouse spleen. Nucl, nucleus. (b) Immunohistochemistry staining of Sox2 and MPO
Supplemental Material
Supplemental Material 1 Figure S1. Phylogenetic analysis of Cep72 and Lrrc36, comparative localization of Cep72 and Lrrc36 and Cep72 antibody characterization (A) Phylogenetic alignment of Cep72 and Lrrc36
Supplementary Methods
Supplementary Methods Reverse transcribed Quantitative PCR. Total RNA was isolated from bone marrow derived macrophages using RNeasy Mini Kit (Qiagen), DNase-treated (Promega RQ1), and reverse transcribed
Supporting information
Supporting information Construction of strains and plasmids To create ptc67, a PCR product obtained with primers cc2570-162f (gcatgggcaagcttgaggacggcgtcatgt) and cc2570+512f (gaggccgtggtaccatagaggcgggcg),
Four different active promoter genes were chosen, ATXN7L2, PSRC1, CELSR2 and
SUPPLEMENTARY MATERIALS AND METHODS Chromatin Immunoprecipitation for qpcr analysis Four different active promoter genes were chosen, ATXN7L2, PSRC1, CELSR2 and IL24, all located on chromosome 1. Primer
Supplementary Figure 1. Immunoprecipitation of synthetic SUMOm-remnant peptides using UMO monoclonal antibody. (a) LC-MS analyses of tryptic
Supplementary Figure 1. Immunoprecipitation of synthetic SUMOm-remnant peptides using UMO 1-7-7 monoclonal antibody. (a) LC-MS analyses of tryptic digest from HEK293 cells spiked with 6 SUMOmremnant peptides
AD BD TOC1. Supplementary Figure 1: Yeast two-hybrid assays showing the interaction between
AD X BD TOC1 AD BD X PIFΔAD PIF TOC1 TOC1 PIFΔAD PIF N TOC1 TOC1 C1 PIFΔAD PIF C1 TOC1 TOC1 C PIFΔAD PIF C TOC1 Supplementary Figure 1: Yeast two-hybrid assays showing the interaction between PIF and TOC1
monoclonal antibody. (a) The specificity of the anti-rhbdd1 monoclonal antibody was examined in
Supplementary information Supplementary figures Supplementary Figure 1 Determination of the s pecificity of in-house anti-rhbdd1 mouse monoclonal antibody. (a) The specificity of the anti-rhbdd1 monoclonal
Supplementary Fig. S1. SAMHD1c has a more potent dntpase activity than. SAMHD1c. Purified recombinant SAMHD1c and SAMHD1c proteins (with
Supplementary Fig. S1. SAMHD1c has a more potent dntpase activity than SAMHD1c. Purified recombinant SAMHD1c and SAMHD1c proteins (with concentration of 800nM) were incubated with 1mM dgtp for the indicated
Figure S1. Specificity of immunofluorescence staining in STC-1 cells. STC-1 cells
Supplementary Figures and Tables Figure S1. Figure S1. Specificity of immunofluorescence staining in STC-1 cells. STC-1 cells were treated with donkey-anti rabbit antibody conjugated with Dylight488 or
hnrnp D/AUF1 Rabbit IgG hnrnp M
Mouse IgG Goat IgG Rabbit IgG Mouse IgG hnrnp F Goat IgG Mouse IgG Kb 6 4 3 2 15 5 Supplementary Figure S1. In vivo binding of TERRA-bound RBPs to target RNAs. Immunoprecipitation (IP) assay using 3 mg
Aoki et al.,
JCB: SUPPLEMENTAL MATERIAL Aoki et al., http://www.jcb.org/cgi/content/full/jcb.200609017/dc1 Supplemental materials and methods Calculation of the raw number of translocated proteins First, the average
SUPPLEMENTARY INFORMATION
DOI: 10.1038/ncb2271 Supplementary Figure a! WM266.4 mock WM266.4 #7 sirna WM266.4 #10 sirna SKMEL28 mock SKMEL28 #7 sirna SKMEL28 #10 sirna WM1361 mock WM1361 #7 sirna WM1361 #10 sirna 9 WM266. WM136
DOI: 10.1038/ncb3259 A Ismail et al. Supplementary Figure 1 B 60000 45000 SSC 30000 15000 Live cells 0 0 15000 30000 45000 60000 FSC- PARR 60000 45000 PARR Width 30000 FSC- 15000 Single cells 0 0 15000
PHT1;2-CFP YFP-PHF + PHT1;2-CFP YFP-PHF
YFP-PHF1 CFP-PHT1;2 PHT1;2-CFP YFP-PHF + PHT1;2-CFP YFP-PHF + CFP-PHT1;2 Negative control!-gfp Supplemental Figure 1: PHT1;2 accumulation is PHF1 dependent. Immunoblot analysis on total protein extract
Genome-wide CRISPR screen reveals novel host factors required for Staphylococcus aureus α-hemolysin-mediated toxicity
Genome-wide CRISPR screen reveals novel host factors required for Staphylococcus aureus α-hemolysin-mediated toxicity Sebastian Virreira Winter, Arturo Zychlinsky and Bart W. Bardoel Department of Cellular
Xu et al., Supplementary Figures 1-7
Xu et al., Supplementary Figures 1-7 Supplementary Figure 1. PIPKI is required for ciliogenesis. (a) PIPKI localizes at the basal body of primary cilium. RPE-1 cells treated with two sirnas targeting to
No wash 2 Washes 2 Days ** ** IgG-bead phagocytosis (%)
Supplementary Figures Supplementary Figure 1. No wash 2 Washes 2 Days Tat Control ** ** 2 4 6 8 IgG-bead phagocytosis (%) Supplementary Figure 1. Reversibility of phagocytosis inhibition by Tat. Human
Description: Nuclear morphology and dynamics in nontargeting sirna transfected cells. HeLa Kyoto
Title of file for HTML: Supplementary Information Description: Supplementary Figures and Supplementary Tables Title of file for HTML: Supplementary Movie 1 Description: Nuclear morphology and dynamics
Sarker et al. Supplementary Material. Subcellular Fractionation
Supplementary Material Subcellular Fractionation Transfected 293T cells were harvested with phosphate buffered saline (PBS) and centrifuged at 2000 rpm (500g) for 3 min. The pellet was washed, re-centrifuged
Fluorescence Light Microscopy for Cell Biology
Fluorescence Light Microscopy for Cell Biology Why use light microscopy? Traditional questions that light microscopy has addressed: Structure within a cell Locations of specific molecules within a cell
Artificial targeting of misfolded cytosolic proteins to endoplasmic reticulum as a mechanism for clearance
Supplementary Information Artificial targeting of misfolded cytosolic proteins to endoplasmic reticulum as a mechanism for clearance Fen Liu 1, Deanna M. Koepp 2, and Kylie J. Walters 1* 1 Protein Processing
Supplementary Fig. 1 Identification of Nedd4 as an IRS-2-associated protein in camp-treated FRTL-5 cells.
Supplementary Fig. 1 Supplementary Fig. 1 Identification of Nedd4 as an IRS-2-associated protein in camp-treated FRTL-5 cells. (a) FRTL-5 cells were treated with 1 mm dibutyryl camp for 24 h, and the lysates
Hossain_Supplemental Figure 1
Hossain_Supplemental Figure 1 GFP-PACT GFP-PACT Motif I GFP-PACT Motif II A. MG132 (1µM) GFP Tubulin GFP-PACT Pericentrin GFP-PACT GFP-PACT Pericentrin Fig. S1. Expression and localization of Orc1 PACT
Chapter 17: Immunization & Immune Testing. 1. Immunization 2. Diagnostic Immunology
Chapter 17: Immunization & Immune Testing 1. Immunization 2. Diagnostic Immunology 1. Immunization Chapter Reading pp. 505-511 What is Immunization? A method of inducing artificial immunity by exposing
1. Immunization. What is Immunization? 12/9/2016. Chapter 17: Immunization & Immune Testing. 1. Immunization 2. Diagnostic Immunology
Chapter 17: Immunization & Immune Testing 1. Immunization 2. Diagnostic Immunology 1. Immunization Chapter Reading pp. 505-511 What is Immunization? A method of inducing artificial immunity by exposing
SUPPLEMENTARY INFORMATION. Supplementary Figures 1-8
SUPPLEMENTARY INFORMATION Supplementary Figures 1-8 Supplementary Figure 1. TFAM residues contacting the DNA minor groove (A) TFAM contacts on nonspecific DNA. Leu58, Ile81, Asn163, Pro178, and Leu182
X2-C/X1-Y X2-C/VCAM-Y. FRET efficiency. Ratio YFP/CFP
FRET efficiency.7.6..4.3.2 X2-C/X1-Y X2-C/VCAM-Y.1 1 2 3 Ratio YFP/CFP Supplemental Data 1. Analysis of / heterodimers in live cells using FRET. FRET saturation curves were obtained using cells transiently
Supplementary material to Alterations in the properties of the cell membrane due to glycosphingolipid accumulation in a model of Gaucher disease
Supplementary material to Alterations in the properties of the cell membrane due to glycosphingolipid accumulation in a model of Gaucher disease Gyula Batta, Lilla Soltész, Tamás Kovács, Tamás Bozó, Zoltán
T H E J O U R N A L O F C E L L B I O L O G Y
T H E J O U R N A L O F C E L L B I O L O G Y Supplemental material Monteiro et al., http://www.jcb.org/cgi/content/full/jcb.201306162/dc1 Figure S1. 3D deconvolution microscopy analysis of WASH and exocyst
Figure S1 early log mid log late log stat.
Figure S1 early log mid log late log stat. C D E F Fig. S1. The chromosome-expressed EI localizes to the cell poles independent of the growth phase and growth media. MG1655 (ptsi-mcherry) cells, which
SUPPLEMENTARY INFORMATION
doi:10.1038/nature10016 Supplementary discussion on binding site density for protein complexes on the surface: The density of biotin sites on the chip is ~10 3 biotin-peg per µm 2. The biotin sites are
Supplemental Figure S1. Nucleotide and deduced amino acid sequences of pepper CaHSP70a (Capsicum annuum heat shock protein 70a) cdna.
Supplemental Figure S1. Nucleotide and deduced amino acid sequences of pepper (Capsicum annuum heat shock protein 70a) cdna. Translation initiation codon is shown in bold typeface; termination codon is
T H E J O U R N A L O F C E L L B I O L O G Y
Supplemental material Kunduri et al., http://www.jcb.org/cgi/content/full/jcb.201405020/dc1 T H E J O U R N A L O F C E L L B I O L O G Y Figure S1. Subcellular localization and interactions of PAPLA1.
Nature Structural & Molecular Biology: doi: /nsmb.3308
Supplementary Figure 1 Analysis of CED-3 autoactivation and CED-4-induced CED-3 activation. (a) Repeat experiments of Fig. 1a. (b) Repeat experiments of Fig. 1b. (c) Quantitative analysis of three independent
Antibodies and reagents Construction and characterization of YFP-TF chimera
Antibodies and reagents Antibodies: mouse monoclonal antibodies (mab) against PDI (clone 34, BD Transduction and RL90, Affinity Bioreagents); rabbit polyclonals against bovine PDI (SPA-890, Stressgen and
Supplementary Figure S1. Binding of HSA mutants to hfcrn. (a) The levels of titrated amounts of HSA
Supplementary Figure S1. Binding of HSA mutants to hfcrn. (a) The levels of titrated amounts of HSA variants (5.0-0.002 μg/ml) directly coated in the wells at ph 6.0 were controlled using a horseradish
i-stop codon positions in the mcherry gene
Supplementary Figure 1 i-stop codon positions in the mcherry gene The grnas (green) that can potentially generate stop codons from Trp (63 th and 98 th aa, upper panel) and Gln (47 th and 114 th aa, bottom
Practice Problems 5. Location of LSA-GFP fluorescence
Life Sciences 1a Practice Problems 5 1. Soluble proteins that are normally secreted from the cell into the extracellular environment must go through a series of steps referred to as the secretory pathway.
Supplemental data. Supplemental Materials and Methods
Supplemental data Supplemental Materials and Methods Transfection of plasmid. Transfection of plasmids into FRTL5 cells was performed using Lipofectamine LTX with Plus reagent (Invitrogen) according to
Supplementary Information. A superfolding Spinach2 reveals the dynamic nature of. trinucleotide repeat RNA
Supplementary Information A superfolding Spinach2 reveals the dynamic nature of trinucleotide repeat RNA Rita L. Strack 1, Matthew D. Disney 2 & Samie R. Jaffrey 1 1 Department of Pharmacology, Weill Medical
SUPPLEMENTARY INFORMATION FILE
SUPPLEMENTARY INFORMATION FILE Existence of a microrna pathway in anucleate platelets Patricia Landry, Isabelle Plante, Dominique L. Ouellet, Marjorie P. Perron, Guy Rousseau & Patrick Provost 1. SUPPLEMENTARY
transcription and the promoter occupancy of Smad proteins. (A) HepG2 cells were co-transfected with the wwp-luc reporter, and FLAG-tagged FHL1,
Supplementary Data Supplementary Figure Legends Supplementary Figure 1 FHL-mediated TGFβ-responsive reporter transcription and the promoter occupancy of Smad proteins. (A) HepG2 cells were co-transfected
JCB. Supplemental material THE JOURNAL OF CELL BIOLOGY. Kimura et al.,
Supplemental material JCB Kimura et al., http://www.jcb.org/cgi/content/full/jcb.201503023/dc1 THE JOURNAL OF CELL BIOLOGY Figure S1. TRIMs regulate IFN-γ induced autophagy. (A and B) HC image analysis
HEK293T. Fig. 1 in the
Supplementary Information Supplementary Figure 1 Zinc uptake assay of hzip4 and hzip4-δecd transiently expressed in HEK293T cells. The results of one representative e experiment are shown in Fig. 1 in
Supplementary Information to: Genome-wide Real-time in vivo Transcriptional Dynamics During Plasmodium falciparum. Blood-stage Development
Supplementary Information to: Genome-wide Real-time in vivo Transcriptional Dynamics During Plasmodium falciparum Blood-stage Development Painter et al. 1 of 8 Supplementary Figure 1: Supplementary Figure
supplementary information
DOI: 10.1038/ncb1977 Figure S1 a. Immunofluorescence analysis of IFT20 localization in PBL costained with anti-β-tubulin antibodies. b. Immunofluorescence analysis of IFT20 localization in Jurkat cells,
fibrils, however, oligomeric structures and amorphous protein aggregates were
Supplementary Figure 1: Effect of equimolar EGCG on αs and Aβ aggregate formation. A D B E αs C F Figure S1: (A-C) Analysis of EGCG treated αs (1 µm) aggregation reactions by EM. A 1:1 molar ratio of EGCG
Supplementary Figure 1 Phosphorylated tau accumulates in Nrf2 (-/-) mice. Hippocampal tissues obtained from Nrf2 (-/-) (10 months old, 4 male; 2
Supplementary Figure 1 Phosphorylated tau accumulates in Nrf2 (-/-) mice. Hippocampal tissues obtained from Nrf2 (-/-) (10 months old, 4 male; 2 female) or wild-type (5 months old, 1 male; 11 months old,
SUPPLEMENTARY INFORMATION
(Supplementary Methods and Materials) GST pull-down assay GST-fusion proteins Fe65 365-533, and Fe65 538-700 were expressed in BL21 bacterial cells and purified with glutathione-agarose beads (Sigma).
Actin cap associated focal adhesions and their distinct role in cellular mechanosensing
Actin cap associated focal adhesions and their distinct role in cellular mechanosensing Dong-Hwee Kim 1,2, Shyam B. Khatau 1,2, Yunfeng Feng 1,3, Sam Walcott 1,4, Sean X. Sun 1,2,4, Gregory D. Longmore
Supplementary information; Mungamuri et al., 2006
Supplementary information; Mungamuri et al., 6 Antibodies used for western blotting: The following antibodies were used for western blotting: antiser473 Akt (#4), antiakt (#97), antiser9 Gsk 3b (#9336),
SAS6-like protein in Plasmodium indicates that conoid-associated apical complex proteins persist in invasive stages within the mosquito vector
SAS6-like protein in Plasmodium indicates that conoid-associated apical complex proteins persist in invasive stages within the mosquito vector Richard J. Wall 1,#, Magali Roques 1,+, Nicholas J. Katris
Phenotypic lentivirus screens to identify functional single domain antibodies
ARTICLE NUMBER: 16080 DOI: 10.1038/NMICROBIOL.2016.80 Phenotypic lentivirus screens to identify functional single domain antibodies Florian I. Schmidt, Leo Hanke, Benjamin Morin, Rebeccah Brewer, Vesna
Supplementatry Fig 1. Domain structure, biophysical characterisation and electron microscopy of a TD. (a) XTACC3/Maskin and XMAP215/chTOG domain
Supplementatry Fig 1. Domain structure, biophysical characterisation and electron microscopy of a TD. (a) XTACC3/Maskin and XMAP215/chTOG domain architecture. Various C-terminal fragments were cloned and
Supplementary Information Cyclin Y inhibits plasticity-induced AMPA receptor exocytosis and LTP
Supplementary Information Cyclin Y inhibits plasticity-induced AMPA receptor exocytosis and LTP Eunsil Cho, Dong-Hyun Kim, Young-Na Hur, Daniel J. Whitcomb, Philip Regan, Jung-Hwa Hong, Hanna Kim, Young
Supplementary Figure S1. Immunodetection of full-length XA21 and the XA21 C-terminal cleavage product.
Supplementary Information Supplementary Figure S1. Immunodetection of full-length XA21 and the XA21 C-terminal cleavage product. Total protein extracted from Kitaake wild type and rice plants carrying