Nanog-Luc. R-Luc. 40 HA-Klf Relative protein level of Klf USP21. Vector USP2 USP21 *** *** *** ***
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1 :Flag Relative protein level of Relative protein level of : Flag Vec 21LV 21SV 2 : HA HA- HA- HA- HA-Klf4 Relative mrna expression Relative protein level of Relative protein level of Relative protein level of Klf4 -Report er Relative Activity a pcdna3.1-nanog-f-luc-ires-r-luc b CMV F-Luc IRES R-LUC -Luc m NES LV C H D 566 AA CMV DUB SV C H D 479 AA R-Luc c e Relative DUBs stabilize ability = F-Luc / R-Luc CHX(h) CHX(h) CHX(h) HA- 35 HA- HA-Klf Time after CHX(h) f Time after CHX(h) g Time after CHX(h) d CA - - i 20 5 Flag- HA- MEF WT KO CHX(h) j FL A B HA- HA- WT KO Time after CHX(h) : Flag UCH - FL A B Binding - - FL A B USP2 CHX(h) HA- Flag-USP Time after CHX(h) k USP2 N H C1 W C2 h Usp21 Binding - - Flag-USP HA Flag - /- /- HA Flag Usp2 HA- shcon sh shusp2 Flag HA Flag HA HA- Flag HA- Flag- HA- Flag-
2 Supplementary Figure 1. directly stabilizes and interacts with. (a). A schematic represents screening model for deubiquitinase using dual luciferase reporter system. (b). Schemes of LV and SV. The alternatively spliced sequence in SV is indicated with dashed line. (c). The half-life of stem cell transcriptional factors (,, Klf4). Statistical analysis was presented in the lower panel. Data are means±s.d. (n=3). (d). was co-transfected with WT or C221A together with -luciferase reporter. The firefly luciferase activity was measured and normalized to the renilla luciferase activity in the same sample and then normalized to vector control. Data are means±s.d. (n=3). P<5, P<01 vs (-) and () condition (Student s t-test). (e). WT and knockout MEFs were infected with virus carrying Flag- and then treated with CHX ( µg ml -1 ) for indicated time intervals, protein levels of and were analyzed by western blotting. Data are means±s.d. (n=3), P< 5 vs WT (Student s t- test). (f). The half-life of upon overexpression of and USP2, respectively. Statistical analysis of was presented in the lower panel. Data are means±s.d. (n=3), P< 01 vs (Student s t-test). (g). The relative knockdown efficiency of and USP2 were detected by Real-time PCR. Data are means±s.d. (n=3), P< 1,P< 01 vs shcon (Student s t-test). (h). Flag-USP2 and was coexpressed with HA- in HEK293T cells. The associated and USP2 or were analyzed by Western blotting. (i). The co- assay in HEK293T cells revealed that HA- could interact with instead of or or Klf4. (j). Schematic diagram of (top panel). The domain of involved in its interaction with. (k). Schematic diagram showing the structure of and its deletion constructs (left). N, interference domain; H, DNA-binding homeodomain; C (C1WC2), activation domain. The domain of involved in its interaction with..
3 IB: HA : Flag (Ub) n WT WT K138R K169R -Reporter Relative Activity K138R/K169R 16KR a His-Ub HA- Flag-USP HA- Flag-USP Ni-NTA b - c 5 0 NS NS - WT KR WT KR WT KR 50ng 0ng 0ng Supplementary Figure 2. deubiquitinates. (a). HA- and His-ubiquitin were coexpressed with USP2 or in HEK293T cells. After MG132 ( μm) treatment for 6 hrs, the ubiquitinated proteins were pulled down under denaturing conditions using Ni-NTA agarose beads and the ubiquitination of was detected by western blotting using an anti-ha antibody. (b). WT or lysine-mutated and were coexpressed in HEK293T cells. After treatment with MG132 ( μm) for 6 hrs, was immunoprecipitated with an anti-flag antibody, and the associated and were analyzed by western blotting using either an anti- or Flag antibody. (c). WT or lysine-mutated was co-transfected with activity luciferase reporter gene into HEK293T cells for 24 hrs. The firefly luciferase activity was measured and normalized to the renilla luciferase activity in the same sample and then normalized to vector control. Data are means±s.d. (n=3). P<5 (Student s t-test).
4 AP colonies Relative expression GFP colonies Relative Expression AP colonies Relative Expression a Usp21 Floxp Allele 5 7 Genotyping b EV Cre Cre c shrna NCU1U3 d shnc sh-1 sh-3 sh Qct4 Rex1 Fgf5 Nestin Desmin Gata4 Sox17 e LIF(-) CTL -SV-1 -SV-2 -LV-1 -LV-2 g Phase N= OE OE fem.intern 7 6 AP f ectoderm mesoderm endoderm EV SV-1 SV-2 LV-1 LV-2 h WT KO WT KO i OSKN OSKN 21SV OSKN 21LV OSKN 21CA WT KO WT KO 0 OSKN - SV LV CA
5 Supplementary Figure 3. is required for mesc self-renewal. (a). PCR was used to detect Usp21 Loxp/Loxp ESCs. (b). Western blotting was used to detect Cre-induced loxp recombination in Usp21 Loxp/Loxp ESCs infected with Cre lentivirus. The anti- antibody is from abgent. (c). Statistic of analysis of Figure 3B. Data are means±s.d. (n=3). P< 01 vs shnc (Student s t- test). (d). Real-time PCR analysis of self-renewal and lineage markers upon E14 cells infected with virus carrying GFP together with control shrna or shrna targeting or. Ectoderm (ECTO), endoderm (ENDO), mesoderm (MESO). Data are means±s.d. (n=3). P<1, P<01 vs shnc (Student s t-test). (e). Morphology and AP staining of E14 cells stably expressing Flag- SV/LV cultured in LIF-free condition for 3 days. Scale bar, 0 µm. (f). The protein levels of,, and in E14 cells stably expressing were analyzed by western blotting. Real-time PCR analysis of self-renewal markers in E14 cells stably expressing. Data are means±s.d. (n=3). P<5, P<1, P<01 vs EV (Student s t-test). (g). Teratoma formation from E14 cells infected with vector or. Scale bar, 0 μm. (h). AP intensity and colonies assessed at stages (day 12) of ips induction in WT and knockout MEF cells. Mean values±s.d. of a representative experiment measured in triplicate are shown. Scale bar, 1 mm. P<5 vs WT (Student s t-test). knockout MEF cells were tested by Real-time PCR and immunoblotting analysis. The anti- antibody is from Abgent. (i). GFP colonies assessed at stages (day 16) of ipsc induction in GFP reporter MEF cell. Mean values±s.d. of a representative experiment measured in triplicate are shown. Scale bar, 1 mm. P<1 vs OSKN condition (Student s t-test).
6 Relative mrna expression DMSO Cryptotanshinone Relative Expression a Rex1 Usp21SV Usp21LV RA Day0 Day1 Day2 Day3 Day4 Day5 b Day c p-stat3(y5) STAT3 Usp21LV Usp21SV Usp2 DMSO Cryptotanshinone d e -LIF(d) p-stat3(y5) STAT3 p-stat3(y5) RA(d) STAT3 Supplementary Figure 4. is a transcriptional target of LIF/STAT3. (a and b). Real-time PCR (a) and immunoblot (b) analyses of pluripotency markers and in E14 cells cultured in RA containing medium for 5 days. Data are means±s.d. (n=3). P<5, P<1, P<01 vs Day0 (Student s t-test). was detected by an anti- from Abgent. (c). Cryptotanshinone ( μm) inhibited the expression of. The expression of was detected by western blot and realtime PCR after treated with cryptotanshinone for 8 hrs. Data are means±s.d. (n=3). P<01 vs DMSO (Student s t-test). Anti- antibody is from abgent. (d and e). The protein level of p-stat3 decreased significantly by LIF withdraw (d) and RA treatment (e) in E14 cells.
7 Normal Gel : Flag p-tag Gel : Flag :Flag :Flag a b c HA- - EGF - - SCH HA- HA- HA-1 HA- MEK1 CA HA- HA (Ub) n Ni-NTA HA- - - HA Flag- His-Ub - MEK1 CA Flag- 1 HA d HA-1 HA-1 HA-1 - LV SV e LV HA MEK1 CA p- HA-1 SV - f WTSAWT HA-1 - MEK1 CA - p- p- Supplementary Fig 5. is phosphorylated by 1. (a). Transfected HEK293T cells were treated with EGF (0 ng ml -1 ) for min and SCH ( µm) for 4 hrs. were immunoprecipitated with an anti-flag antibody, and the associated was analyzed by western blotting using HA antibody. (b). Phosphorylation blocked the interaction between and. (c). Phosphorylation blocks mediated deubiquitination. The transfected HEK293T cells were treated with MG132 (μm) for 6 hrs, the ubiquitinated proteins were pulled down under denaturing and analyzed by western. (d). Binding of 1 to. (e). HEK293T cells were transfected with indicated combination of, 1 and MEK1CA plasmids. Phos-Tag SDS-PAGE was applied to detect the band shift of caused by phosphorylation. LV/SV and HA- 1 were determined by regular SDS-PAGE with the anti-flag or anti-ha antibodies, respectively. (f). The specificity of antibody against the phosphorylaed at S539.
8 Relative Expression :HA WT WT S93D S335D S539D DMSO PD03901 SCH a b c CHX(h) HA- Ctl WT SA HA- HA- HA- - (abcam) 35 d 6 e 4 2 WT SA SD WT SD SA 0 OCT4 Rex1 Fgf5 Nestin Demin Gata4 Sox17 Supplementary Figure 6. Phosphorylation of at 539 blocks its effect on. (a). The effect of WT and S539A on the stability of HA- HEK293T cells. (b). S539D but not S93D and S335D blocked the binding of to. (c). Inhibition of stabilized. E14 cells were cultured with LIF-free condition with the MEK inhibitor PD03901 (1 μm) or 1/2 inhibitor SCH ( μm). Media were changed every 24 hrs over 4 days. The anti- antibody is from Abcam. (d). The effect of and its mutants on the gene expression in E14 cells. E14 cells stably expressing (WT, S539A, S539D) were cultured in N2B27 medium overnight and treated with mfgf4 ( ng ml -1 ) for 12 hrs. Data are means±s.d. (n=3). P<1, P<01 vs (two-way ANOVA test); P< 5, P< 1, P< 01 vs WT (two-way ANOVA test). (e). Tumor images were taken 5 weeks after inoculation.
9 LV Normalized to H3 LV SV SV a Merge DAPI b (abcam) ubh2a(k119) H3K4me3 H3K27me3 d RA(d) H2A H3 ubh2a H2A H3K4me3 H3 - WT SD SA e c Rosa26 locus Donor Plasmid Exon1 Homologous arm Exon1 H3K4me3 Ch sgrna Cas9 CAG-Flag- CAG-Flag- Sall4 Lefty1 Cdx2 Hoxa Exon2 Homologous arm Exon2 Control WT SD SA Supplementary Figure 7. is recruited to gene promoters and modulates H2A ubiquitination. (a). was re-localized to the nucleus by coexpressed. HA- was coexpressed with LV/SV in HeLa cells. The subcellular location of (green) and (red) was examined by immunofluorescent staining (nuclei were stained with DAPI; blue). (b). The effect of RA treatment on histone ubiquitination and methylation in E14. The anti- antibody is from abcam. (c). Schematic overview of strategy to generate (WT, S539A, S539D) knock-in E14 stable cell line. (d). The effect of and its mutants on histone ubiquitination and methylation in E14 cells. (e). The effect of and its mutants on the level of H3K4me3 at regulated gene promoters in E14 cells. Data are means±s.d. (n=3). P<1, P<01 vs WT (two-way ANOVA test); P< 01 vs Control (two-way ANOVA test).
10 GST- His- Fig. 1b Fig. 1e HA- Fig. 1g Fig. 1c HA- Fig. 1h Fig. 1d Klf4 HA- HA- Fig. 1i His- Fig. 1j Fig. 1k Klf4 Suppl. Fig. 1i HA-// /Klf4 Suppl. Fig. 1c Suppl. Fig. 1d Suppl. Fig. 1f HA- HA- HA-Klf4 Suppl. Fig. 1e Flag- HA- Suppl. Fig. 1h HA- /2 Flaf-LV/SV/2 HA- Suppl. Fig. 1j Suppl. Fig. 1k HA- HA- Flag- Supplementary Figure 8. Uncropped scans of blots. Uncropped scans of Figure 1 and Supplementary Figure 1.
11 (Ub)n (Ub)n (Ub)n (Ub)n (Ub)n Fig. 2a Fig. 2b Fig. 2c Fig. 2e Flag- HA- Flag- His- Suppl. Fig. 2a Fig. 2f Suppl. Fig. 2b Suppl. Fig. 2c HA- Flag-USP Suppl. Fig. 3b Suppl. Fig. 3f Suppl. Fig. 3h Fig. 4b Cre Fig. 4d HA-STAT3 Fig. 4e HA-STAT3 Fig. 4f STAT3 Fig. 4g Flag-STAT3 Suppl. Fig. 4b Suppl. Fig. 4c Suppl. Fig. 4d Suppl. Fig. 4e p-stat3 STAT3 p-stat3 STAT3 p-stat3 STAT3 Supplementary Figure 9. Uncropped scans of blots. Uncropped scans of Figure 2-4 and Supplementary Figure 2-4.
12 (Ub)n (Ub)n Fig. 5a Fig. 5d Fig. 5h Fig. 6b HA- Suppl. Fig. 6c 1/2 phosphosubstrate p- Suppl. Fig. 5e p- Fig. 5i Fig. 6c Fig. 7a Fig. 5b phos-tag gel HA- ubh2a H2A H3K4me3 H3K27me3 H3 Ub p- Fig. 5f GST-1 GST HA-1 Fig. 6d Fig. 7b ubh2a H2A H3K4me3 H3 Fig. 6a Suppl. Fig. 7b Suppl. Fig. 6a ubh2a H2A H3K4me3 H3K27me3 H3 Supplementary Figure. Uncropped scans of blots. Uncropped scans of Figure 5-7 and Supplementary Figure 5-7. Suppl. Fig. 5a Suppl. Fig. 5c 1/2 phospho- Substrate Ab HA- Suppl. Fig. 5f p- Ub phos-tag Gel Suppl. Fig. 5b Fig. 5c Flag- HA- Fig. 5g Suppl. Fig. 6b Suppl. Fig. 5d p- HA- HA-1 HA- Suppl. Fig. 7d HA- HA- ubh2a H2A H3K4me3 H3 p-
13 Supplementary Table 1 Gene Expression Primers Gene Expression primers NAME 5 Fwd 3 Rev CTCAAGTCCTGAGGCTGACA TGAAACCTGTCCTTGAGTGC TAGGTGAGCCGTCTTTCCAC GCTTAGCCAGGTTCGAGGAT Rex1 GACGGATACCTAGAGTGCATCA GAAGGGAACTCGCTTCCAGAA AGGGCTGGGAGAAAGAAGAG CCGCGATTGTTGTGATTAGT SV GATGAAAGGCTCAAGAAACTGGAG TGCTGCTCAAACACTGTAGC LV CATGGCTCCTTCCACATGAT AGGGCACCAATCACATCTGC Fgf5 GAAATATTTGCTGTGTCTCAGGG TAAATTTGGCACTTGCATGG Nestin AGAAGACGGTGTGTCTGCTT TAGAGGTGCAGCAGCTGCAG Demin TGACAACCTGATAGACGACC TTAAGGAACGCGATCTCCTC Gata4 TCCTACTCCAGCCCCTACC GTAGTGTCCCGTCCCATCTC Sox17 GAACAGTTGAGGGGCTACAC GTTTAGGGTTTCTTAGATGC USP2 CCATGGGAGGCCACTATACAGCCTA CAAATAGGCGTCGCTGGT
14 Supplementay Table 2. Primers for Ch-qPCR Ch-qPCR primers NAME 5 Fwd 3 Rev AGTGCCAGCCTCGTCCCGTAGA CAAAATG AAGTGGGCCCCGGC CTTCTCCAT Sall4 TGGACAACCCAGAGCTGAAT CAGGAGGGGCTGTCCTATTT CCAAGACTGGAGCTCACAATC CAGGTGGAGCCTGAAAAGAAG Lefty1 GTCCAGACAGGCTTTTGTGT AGTCTGCGGAGGAATGGTA Cdx2 GGACTCCGCGAGCCAA CTCAGCCCACGGTGCTC Hoxa CTGGCTCTTGAACCTGTACCCC CAAGGGTGCTTCCAAATAGTC SupplementaryTable 3. binding sites at genomic locations Genesymbol TSS TTS Refseq ID chromsome Sall NM_ chr NM_ chr3 Lefty NM_0094 chr1 Cdx NM_ chr5 Hoxa NM_ chr6
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monoclonal antibody. (a) The specificity of the anti-rhbdd1 monoclonal antibody was examined in
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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
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SUPPLEMENTRY INFORMTION DOI:.38/ncb Kdmb locus kb Long isoform Short isoform Long isoform Jmj XX PHD F-box LRR Short isoform XX PHD F-box LRR Target Vector 3xFlag Left H Neo Right H TK LoxP LoxP Kdmb Locus
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Supplementary Fig. 1
a FL (1-2266) NL (1-1190) CL (1191-2266) HA-ICE1: - HA-ICE1: - - - FLAG-ICE2: + + + + FLAG-ELL: + + + + + + IP: anti-ha FLAG-ICE2 HA-ICE1-FL HA-ICE1-NL HA-ICE1-CL FLAG-ICE2 b IP: anti-ha FL (1-2266) NL
b alternative classical none
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Cell, Volume 135 Supplemental Data Hypothalamic IKKβ/NF-κB and ER Stress Link Overnutrition to Energy Imbalance and Obesity Xiaoqing Zhang, Guo Zhang, Hai Zhang, Michael Karin, Hua Bai, and Dongsheng Cai
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DOI: 10.1038/ncb1864 Figure S1 Apak specifically inhibits p53 transcriptional activity. Transcription activity of p53 was measured in U2OS (p53 wild-type) and H1299 (p53 deficient) cells which were transfected
Supplementary Figure 1 PARP1 is involved in regulating the stability of mrnas from pro-inflammatory cytokine/chemokine mediators.
Supplementary Figure 1 PARP1 is involved in regulating the stability of mrnas from pro-inflammatory cytokine/chemokine mediators. (a) A graphic depiction of the approach to determining the stability of
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SUPPLEMENTARY INFORMATION Supplementary figures Supplementary Figure 1: Suv39h1, but not Suv39h2, promotes HP1α sumoylation in vivo. In vivo HP1α sumoylation assay. Top: experimental scheme. Middle: we
(a) Scheme depicting the strategy used to generate the ko and conditional alleles. (b) RT-PCR for
Supplementary Figure 1 Generation of Diaph3 ko mice. (a) Scheme depicting the strategy used to generate the ko and conditional alleles. (b) RT-PCR for different regions of Diaph3 mrna from WT, heterozygote
Supplementary Fig. 1 Kinetics of appearence of the faster migrating form of Bcl-10.
α-cd3 + α-cd28: Time (min): + + + + + + + + + 0 5 15 30 60 120 180 240 300 360 360 n.s. Supplementary Fig. 1 Kinetics of appearence of the faster migrating form of. Immunoblot of lysates from Jurkat cells
Supplementary Figure 1.
Supplementary Figure 1. Quantification of western blot analysis of fibroblasts (related to Figure 1) (A-F) Quantification of western blot analysis for control and IR-Mut fibroblasts. Data are expressed
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(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).
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
Schematic representation of the endogenous PALB2 locus and gene-disruption constructs
Supplementary Figures Supplementary Figure 1. Generation of PALB2 -/- and BRCA2 -/- /PALB2 -/- DT40 cells. (A) Schematic representation of the endogenous PALB2 locus and gene-disruption constructs carrying
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DOI: 10.1038/ncb3240 Supplementary Figure 1 GBM cell lines display similar levels of p100 to p52 processing but respond differentially to TWEAK-induced TERT expression according to TERT promoter mutation
Supplementary Information
Supplementary Information Supplementary Figure S1 (a) P-cRAF colocalizes with LC3 puncta. Immunofluorescence (IF) depicting colocalization of P-cRAF (green) and LC3 puncta (red) in NIH/3T3 cells treated
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
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
Supplementary Figure 1. GST pull-down analysis of the interaction of GST-cIAP1 (A, B), GSTcIAP1
Legends Supplementary Figure 1. GST pull-down analysis of the interaction of GST- (A, B), GST mutants (B) or GST- (C) with indicated proteins. A, B, Cell lysate from untransfected HeLa cells were loaded
gacgacgaggagaccaccgctttg aggcacattgaaggtctcaaacatg
Supplementary information Supplementary table 1: primers for cloning and sequencing cloning for E- Ras ggg aat tcc ctt gag ctg ctg ggg aat ggc ttt gcc ggt cta gag tat aaa gga agc ttt gaa tcc Tpbp Oct3/4
HPV E6 oncoprotein targets histone methyltransferases for modulating specific. Chih-Hung Hsu, Kai-Lin Peng, Hua-Ci Jhang, Chia-Hui Lin, Shwu-Yuan Wu,
1 HPV E oncoprotein targets histone methyltransferases for modulating specific gene transcription 3 5 Chih-Hung Hsu, Kai-Lin Peng, Hua-Ci Jhang, Chia-Hui Lin, Shwu-Yuan Wu, Cheng-Ming Chiang, Sheng-Chung
USP19 modulates autophagy and antiviral immune responses by. deubiquitinating Beclin-1
USP19 modulates autophagy and antiviral immune responses by deubiquitinating Beclin-1 Shouheng Jin 1,2,, Shuo Tian 1,, Yamei Chen 1,, Chuanxia Zhang 1,2, Weihong Xie, 1 Xiaojun Xia 3,4, Jun Cui 1,3* &
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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
This is the author's accepted version of the manuscript.
This is the author's accepted version of the manuscript. The definitive version is published in Nature Communications Online Edition: 2015/4/16 (Japan time), doi:10.1038/ncomms7780. The final version published
Nature Structural & Molecular Biology: doi: /nsmb Supplementary Figure 1
Supplementary Figure 1 Endogenous gene tagging to study subcellular localization and chromatin binding. a, b, Schematic of experimental set-up to endogenously tag RNAi factors using the CRISPR Cas9 technology,
Supplementary Figure 1. Expressions of stem cell markers decreased in TRCs on 2D plastic. TRCs were cultured on plastic for 1, 3, 5, or 7 days,
Supplementary Figure 1. Expressions of stem cell markers decreased in TRCs on 2D plastic. TRCs were cultured on plastic for 1, 3, 5, or 7 days, respectively, and their mrnas were quantified by real time
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 S1. USP-46 is expressed in several tissues including the nervous system
Supplemental Figure legends Figure S1. USP-46 is expressed in several tissues including the nervous system Transgenic animals expressing a transcriptional reporter (P::GFP) were imaged using epifluorescence
Nature Methods: doi: /nmeth Supplementary Figure 1. Schematics of SAM, dcas9-suntag and dcas9-vpr.
Supplementary Figure 1 Schematics of SAM, dcas9-suntag and dcas9-vpr. (a) Schematics of SAM. SAM consists of two chimeric proteins, dcas9 fused with VP64 and MS2-coat protein fused with p65 and HSF1 activator
Supplementary Figure 1
Supplementary Figure 1 Supplementary Fig. 1 shrna mediated knockdown of ZRSR2 in K562 and 293T cells. (a) ZRSR2 transcript levels in stably transduced K562 cells were determined using qrt-pcr. GAPDH was
TE5 KYSE510 TE7 KYSE70 KYSE140
TE5 KYSE5 TT KYSE7 KYSE4 Supplementary Figure. Hockey stick plots showing input normalized, rank ordered H3K7ac signals for the candidate SE-associated lncrnas in this study. Rpm Rpm Rpm Chip-seq H3K7ac
supplementary information
DOI: 10.1038/ncb1816 A Gal4 Gal4 B Kinase domain C (1502) N (5111191) Interaction with SIAH1 C SIAH1 SIAH1 C (182) SIAH1 N (82282) RING domain Interaction with Input GST GSTSIAH1 1a a N C Input GST GSTSIAH1
Supplementary Fig. 1. (A) Working model. The pluripotency transcription factor OCT4
SUPPLEMENTARY FIGURE LEGENDS Supplementary Fig. 1. (A) Working model. The pluripotency transcription factor OCT4 directly up-regulates the expression of NIPP1 and CCNF that together inhibit protein phosphatase
Nature Structural & Molecular Biology: doi: /nsmb Supplementary Figure 1.
Supplementary Figure 1. Characterization and high expression of Lnc-β-Catm in liver CSCs. (a) Heatmap of differently expressed lncrnas in Liver CSCs (CD13 + CD133 + ) and non-cscs (CD13 - CD133 - ) according
Nature Immunology: doi: /ni Supplementary Figure 1. Zranb1 gene targeting.
Supplementary Figure 1 Zranb1 gene targeting. (a) Schematic picture of Zranb1 gene targeting using an FRT-LoxP vector, showing the first 6 exons of Zranb1 gene (exons 7-9 are not shown). Targeted mice
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Figure S1: Activation of the ATM pathway by I-PpoI. A. HEK293T cells were either untransfected, vector transfected, transfected with an I-PpoI expression vector, or subjected to 2Gy γ-irradiation. 24 hrs
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doi:.38/nature899 Supplementary Figure Suzuki et al. a c p7 -/- / WT ratio (+)/(-) p7 -/- / WT ratio Log X 3. Fold change by treatment ( (+)/(-)) Log X.5 3-3. -. b Fold change by treatment ( (+)/(-)) 8
Figure S1. Sequence alignments of ATRIP and ATR TopBP1 interacting regions.
A H. sapiens 204 TKLQTS--ERANKLAAPSVSH VSPRKNPSVVIKPEACS-PQFGKTSFPTKESFSANMS LP 259 B. taurus 201 TKLQSS--ERANKLAVPTVSH VSPRKSPSVVIKPEACS-PQFGKPSFPTKESFSANKS LP 257 M. musculus 204 TKSQSN--GRTNKPAAPSVSH
Supplementary Methods Plasmid constructs
Supplementary Methods Plasmid constructs. Mouse cdna encoding SHP-1, amplified from mrna of RAW264.7 macrophages with primer 5'cgtgcctgcccagacaaactgt3' and 5'cggaattcagacgaatgcccagatcacttcc3', was cloned
Description of Supplementary Files. File name: Supplementary Information Description: Supplementary figures and supplementary tables.
Description of Supplementary Files File name: Supplementary Information Description: Supplementary figures and supplementary tables. File name: Supplementary Data 1 Description: Differential expression
Supplementary Information
Supplementary Information Sam68 modulates the promoter specificity of NF-κB and mediates expression of CD25 in activated T cells Kai Fu 1, 6, Xin Sun 1, 6, Wenxin Zheng 1, 6, Eric M. Wier 1, Andrea Hodgson
Supplementary Information
Supplementary Information Supplementary Figure 1. Elevated Smurf1 expression is associated with the progression of colorectal cancer. (a) The tumor microarray analysis with anti-smurf1 antibody. (b) Box
Supplementary Information
Supplementary Information Supplementary Figure 1: Identification of new regulators of MuSC by a proteome-based shrna screen. (a) FACS plots of GFP + and GFP - cells from Pax7 ICN -Z/EG (upper panel) and
Supplemental Material Igreja and Izaurralde 1. CUP promotes deadenylation and inhibits decapping of mrna targets. Catia Igreja and Elisa Izaurralde
Supplemental Material Igreja and Izaurralde 1 CUP promotes deadenylation and inhibits decapping of mrna targets Catia Igreja and Elisa Izaurralde Supplemental Materials and methods Functional assays and
a. Increased active HPSE (50 kda) protein expression upon infection with multiple herpes viruses: bovine
Fold change vs uninfeced MOI 0.1 MOI 1 MOI 0.1 MOI 1 MOI 0.001 MOI 0.01 Fold change vs uninfected a 6 4 2 50 kda HPSE * ** * * BHV PRV HSV-2 333 ** * 0 - + + - + + - + + b 50 kda HPSE 3 2 ** *** mock inf
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
Supplementary Figure 1. Localization of MST1 in RPE cells. Proliferating or ciliated HA- MST1 expressing RPE cells (see Fig. 5b for establishment of
Supplementary Figure 1. Localization of MST1 in RPE cells. Proliferating or ciliated HA- MST1 expressing RPE cells (see Fig. 5b for establishment of the cell line) were immunostained for HA, acetylated
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
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 Figure 1
Supplementary Figure 1 Virus infection induces RNF128 expression. (a,b) RT-PCR analysis of Rnf128 (RNF128) mrna expression in mouse peritoneal macrophages (a) and THP-1 cells (b) upon stimulation with
Supplementary Figure 1. TSA (10 nmol/l), non-class-selective HDAC inhibitor, potentiates
Supplementary Figure 1. TSA (10 nmol/l), non-class-selective HDAC inhibitor, potentiates vascular calcification (VC). (a) Von Kossa staining shows that TSA potentiated the Pi-induced VC. Scale bar, 100
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