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1 1 1 μm c d EGF + TPA + e f Intensity (Hours) Time (Hours) Reltive luciferse ctivity CAMEK1 FRE reporter Figure S1 inhiitor incresed protein expression nd ctivted inhiited trnscriptionl ctivity of responsive elements driven luciferse reporter. () Lystes of 293T cells were trnsfected with wildtype long with different dosges of nd nlyzed y immunolotting. () Rel time PCR trnscript products of ws mesured, nd the reltive fold of induction ws clculted nd compred etween nd treted cells. (c) Serum strved MCF7 cells were treted with EGF for 8 hrs with or without. Cell lystes were sujected to immunolotting with the ntiodies indicted. (d) Serum strved MCF7 cells were treted with 12Otetrdecnoylphorol 13cette (TPA) for 8 hrs with or without. Cell lystes were sujected to immunolotting with the ntiodies indicted. (e) 293T cells were incuted with methionine nd cysteinefree medium overnight. Cells were treted with or without, pulsed with [ 35 S] Met for 3 minutes, nd chsed for the indicted time intervls. Cell lystes were immunoprecipitted with n nti ntiody nd sujected to SDSPAGE nlyses. Gels were fixed, dried, nd sujected to utordiogrphy (with n intensifying screen). (f) Lystes of 293T cells cotrnsfected with mrked plsmids were sujected to luciferse ssys with FOXOresponsive elements driven luciferse reporter. Grph shows the men vlue of the representtive results from three experiments (n=3) with s.d. conducted in duplictes for ech. All the concentrtions nd time for tretment were descried in the methods. 1
2 CAMEK1 : IP: + p(s344) IP: CAMEK1 WT 3A p(s344) Lyste c d Vector CAMEK1 IP: * p(s344) WT 3A CAMEK1: + + IP: S344 IP: S4 IP: HA e f EGF C EGF + N + 12 mins S Time (hours) PARP Tuulin pakt g Serum strved EGF + h 1 1 ßActin In vitro kinse ssy I Coomsie lue stining Figure S2 Identifiction of phosphoryltion sites of y specific phosphontiody trgeting p (S294, S344 nd S4). () Lystes of 293T cells trnsfected with CAMEK1 were sujected to immunoprecipittion nd immunolotting with the ntiodies indicted. () Lystes of 293T cells cotrnsfected or 3A with CA MEK1 were sujected to immunoprecipittion nd immunolotting (nti nd ntip344). (c) Cells treted with PD9859 for 4 hours efore the lystes were extrcted nd sujected to nlysis s descried in () (lck rrow). The str represents nonspecific nd for P ntiody. (d) Lystes of 293 cells cotrnsfected with or 3A nd CAMEK1 were sujected to immunoprecipittion nd immunolotting (IP: ntips344 or S4 nd ntiha; IB: nti). (e) MCF7 cells, fter serum strvtion, were treted with EGF for 12 minutes with or without then cytoplsmic nd nucler frctiontions were nlyzed y immunolotting with indicted ntiodies. (f) MCF7 cells were extrcted t the indicted times fter serum strvtion nd EGF stimultion nd then sujected to immunolotting with the ntiodies indicted. (g) The nd upshift is locked y inhiitor. MCF7 cells were strved overnight nd stimulted with EGF for 3 minutes. Lystes were left untreted or were treted with nd sujected to immunolotting. (h) directly phosphoryltes in vitro. In vitro kinse ssy ws performed y incuting recominnt ctivted 2 with fulllength. 2
3 # 6 # 1 # 14 ( + ) ( ) # 1 # 3 # 9 ( ) ( + ) R 2 =.538 p= Brest cncer tissues Numer: Actin Figure S3 Inverse correltion of nd expression in humn rest cncer ptients. () The IHC levels of nd from consecutive tumor sections (# 6,1 nd 14 represented + nd ; # 1,3 nd 9 represented nd +) of six ptients. The ptient numers in () nd () re the sme. () Lystes from fourteen rest cncer ptients were sujected to western lotting with indicted ntiodies. The liner regression nlysis ws used to nlyze the nd expression in the lystes of rest cncer ptients. 3
4 Cell growth rtio T3V12RAS GFP FOXOWT FOXO3A FOXO3D Time (dys) DN1/2 c sifoxo sifoxo+foxo Tuulin Cell grwoth rtio Actin d CMEK1 + p (344) IP : IP: Vector CMEK1 MyrAKT IKK p (3) p (644) Totl lyste HAMEK1 HAAKT FlgIKK Figure S4 () 3A mutnt, ut not 3D, inhiited cell growth. NIHV12Rs cells were trnsfected with control vector, wildtype (WT), nd 3A nd 3D mutnts. After sorting with GFP mrker for positive trnsfection, GFPpositive cells were sujected to MTT ssy. Grph shows the men vlue of the representtive results from three experiments (n=3) conducted in duplictes for ech (s.d.) () (c) Forced expressing long with sirna restores protein level s well s inhiits cell growth. MDAMB435 cells were trnsfected with control vector (lne 1), DN1 nd DN2 (lne 2), DN1, DN2 nd psuper sirna (lne 3), DN1, DN2 nd psuper sirna nd (lne 4), nd were sujected to () Western lot nd (c) MTT ssy. MTT ssy (c) ws performed using the sme tch of trnsfected cells s () nd leled the sme s shown in () Lne1 to lne 4. Grph shows the men vlue of the representtive results with s.d. from three experiments (n=3) conducted in duplictes for ech. (d) Lystes of 293T cells cotrnsfected with CAMEK1, MyrAKT or IKKβ were immunoprecipitted with ntiody nd immunolotted with the ps3, ps644 nd totl ntiodies. Totl lystes were seprtely y immnolotted with indicted ntiodies. 4
5 Figure 1 Full gel of Figure 1i CAMEK1 1 IB: GFP IB: IB: GFP Shorter exposure CHX: IB: (Hr) CHX: IB: (Hr) Full gel of Figure 1h Full gel of Figure 1k 3BX 2 3BX 1 NIH3T3 VRAS NIH3T3 VRAS NIH3T3 VRAS NIH3T3 VRAS NIH3T3 VRAS 1 IB: IB: IB: IB: Bim IB: p27 IB: IB: Full gel of Figure 3e p53/ p53, / p53/ p53, / Full gel of Figure 2i WT WT 3A WT WT 3A CAMEK1 : CAMEK1 : PD9859 PD9859 PD9859 PD9859 IgG IB: IP: GFP IB: pserine IP: GFP IB: IB: IB: Figure S5 The full pnels of primry gels from figures indicted. 5
6 Figure 4e 1 IB: 3D IB: IB: Tuulin Figure 4f IB: 3D IB: 9 IB: Tuulin Figure 4g Figure 5c MDAMB435 MDAMB435 WT 3A 3D M WT r (K) 3A 3D 1 1 3A MDAMB435 WT 3A 3D Cspse 3 Tuulin IB: IB: Bim Cleved cspse 3 Figure S6. The full pnels of primry gels from figures indicted. 6
Interplay between NS3 protease and human La protein---- by Ray and Das Supplementary fig 1. NS3 pro
Interply etween tese nd humn L protein---- y Ry nd Ds Supplementry fig 1 1 2 3 4 UV crosslinking ssy: α[ 32 P]UTP leled HCV IRES RNA ws UV-crosslinked to incresing concentrtions (0.1, 0.2 nd 0.4µM) in
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DOI: 1.138/nc2274 EpH4 Prentl -/MDCK EpH4 Prentl -/MDCK - FERM- Figure S1 (kd) delferm- (1-438)- c Prentl MDCK -/MDCK deljfr- Input Control IP IP Input Control IP IP d Control Control Figure S1 () Specificity
COS-1 cells transiently transfected with either HA hgr wt, HA hgr S211A or HA hgr S226A
1 SUPPLEMENTRY FIGURES Fig. 1 & Specificity of the nti-p-s211 nd nti-p-s226 ntibodies COS-1 cells trnsiently trnsfected with either H hgr wt, H hgr S211 or H hgr S226 were treted with 1nM Dex for 1 hour.
a ATP release 4h after induction
doi:1.138/nture9413 ATP relese 4h fter induction of poptosis (nm) 5 ATP 4 3 2 1 UV UV + zvad 1μM 3μM 5μM UV + 1μM 3μM 5μM UV + 18AGA 1μM 3μM 5μM UV + FFA HeL monolyer Scrpe Dye trnsfer HeL HeL-Cx43 HeL-Cx43
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SUPPLEMENTARY INFORMATION doi:10.1038/nture11303 c Supplementry Figure 1: Genertion of INCB18424 persistent B/F3 Epor- V617F (EporVF) cells, which re cross resistnt to other inhiitors. : () Nïve EporVF
Gan et al., Supplemental Figure 1
Gn et l., Supplementl Figure IB: DU45 Unp IB: py-68 IB: perk IB: ERK IB: Akt Unp DU45 DU45 ErB2 - EGF - EGF ErB2 75 75 Supplementl Figure. DU45 nd ells predominntly express nd re highly responsive to EGF.
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doi: 1.138/nture77 c 2 2 1 15 1 1 5 1 1 5 5 129Sv 1.5 h IL-6 / HPRT 3 h 5 h 6 4 2 d 129Sv 4 4 3 3 2 1 1 8 6 4 2 e f g C57/BL6 Poly(dA-dT) Cell numer (normlized to medium control) 1 5 1 5 1 5 Fluorescence
Fluorescence Intensities of. GFP-PAC-1 Strains
DOI: 10.1038/ncb3168 Arbitrry Fluorescence Units 2500 2000 1500 1000 500 0 full length (1-4) Fluorescence Intensities of GFP-PAC-1 Strins ΔPH 392-838 575-4 GFP-PAC-1 Strins 2-610 1-574 b control c pc-1(3
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NCS (ng/ml) Time (min) kd 500 500 0 30 0 30 IP: IP: 112 105 75 IB: ps407 IB: Mdm2 NCS + + IB: IB: tuulin IP input sup NCS (ng/ml) 50 100 500 Time (min) 0 15 30 60 120 15 30 60 120 15 30 60 120 IB: ps407
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DOI: 1.138/nc3386 β-ctin (S) (A) - - - TGN46 116 kd 97 45 level (% of ) 1 8 6 4 2 EEA1 (S) (A) c 1 Reltive mrna levels normlized to β-ctin 8 6 4 2 TFEB KD MCOLN1 KD PI4KIIIβ KD PIP5K1α KD PIP5K1β KD VPS
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Supplementry Figures nd Legends S1 Figure S1. Trgeted FRET sensors of Auror B kinse ctvity., Imges of cells expressing the untrgeted (i), centromere trgeted (ii), or chromtin-trgeted sensors (iii) in mitosis.
Jay S Desgrosellier, Leo A Barnes, David J Shields, Miller Huang, Steven K Lau, Nicolas Prévost, David Tarin, Sanford J Shattil and David A Cheresh
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SUPPLEMENTARY INFORMATION doi:10.1038/nture10177 MDYKDHDGDYKDHDIDYKDD DDKMAPKKKRKVGIHGVPAA MAERPFQCRICMRKFAQSGD LTRHTKIHTGEKPFQCRICM RNFSRSDVLSEHIRTHTGEK PFACDICGKKFADRSNRIKH TKIHTGSQKPFQCRICMRNF SRSDNLSEHIRTHTGEKPFA
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doi:10.1038/nture12040 + + + Glc Gln Supplementry Figure 1., Reltive prolifertion of PDAC cell lines (8988T, Tu8902, Pnc1, Mipc2, PL45 nd MPnc96) nd low pssge primry humn PDAC cell lines (#1 nd #2) under
Figure S1 Yoo et al.
doi:.38/nture6543 8 8 6 6 4 4 d Protoplsts Leves Reltive promoter ctivity (%) Reltive trnscript level 2 2 88 66 44 22 32 2 2 MKK-MYC MPK ctivity nti-mpk6 c ctr MKK - 4 5 4 5 MKK-MYC MPK3 ctivity MPK6 ctivity
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DOI: 1.138/n74 In the formt provided y the uthors nd unedited. d 331p 637p 9p 394p 48p 467p 22p 23p 489p 419p 3p 493p 332p 53p 39p 1 G1: 16.4% S: 73.1% G2/M: 1.5% 2 1 2 E-KO G1: 2.2% S: 7.7% G2/M: 9.1%
Supplementary Figure 1. A TRPC5-like channel in the neurites and somata of aortic baroreceptor neurons.
Supplementl Figure 1 c d e f Supplementry Figure 1. A TRPC5-like chnnel in the neurites nd somt of ortic roreceptor neurons. Cell-ttched ptch recordings from the neurite terminls (-c) nd the somt (d-f)
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SUPPLEMENTARY INFORMATION doi:1.138/nture1371 As Brc11 (null) Brc5-13cK (conditioned llele) Reltive levels of mrna d C 1.2 1.8.6.4.2 Cereellum Ec Ev Ev As KO c Frequency Thymus KO KO 1 neo 11 Ec As 4 loxp
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DOI: 10.1038/nc2473 AP-2 EEA1 APPL1 EEA1 APPL1 EEA1 APPL1 c d e AP-2 FCHO2 merge f g h i k FCHO1 EFC domin FCHO2 EFC domin 10X 1X 3X 10X 10X 1X 3X 10X FCHO1/FCHO2 _ + + + _ + + + PtdIns(4,5)P 2 S P S P
Supplementary Figure 1
Supplementry Figure 1 d6 d8 d9 SSC.575 27.4 35.1 d1 d12 d2 39.6 5.4 67.3 NKX2-5-GFP Supplementry Figure 1 Differentition kinetics of the NKX2-5-GFP HES3 hesc line (Elliott et l., 211). Flow cytometric
WesternBright TM MCF and MCF-IR
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Supplementary information
PP GC B Epithelil cell Peyer s ptch B lymphocyte Dendritic cell Bsophil Neutrophil Mst cell Nturl killer cell T lymphocyte Supplementry informtion EAF2 medites germinl center B cell poptosis to suppress
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doi:1.138/nture9816 control.1 µg/µl.1 µg/µl 1 µg/µl NH2 signl 2xFLAG PS Fc-tg CD4L [116-261] 55 274 146 c counts Stimultion of Rji cells Lysis nd 1 st IP with M2-eds (4 C) PreScission nd FLAG-peptide tretment
Supplemental Figure S1
Supplementl Figure S1 TG nrt1.5- Li et l., 1 nrt1.5- Lin et l., 8 F L CTGCCT R T 5'UTR 3'UTR 1 3 81p (k) nrt1.5- C nrt1.5- Supplementl Figure S1. Phenotypes of the T-DN insertion mutnts (this pper), nrt1.5-
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S shrna S Viility, % of NT sirna trnsfete ells 1 1 Srmle NT sirna Csp-8 sirna RIP1 Atin L929 shrna sirna: Csp8 Atin L929: shrna Csp8 e Viility, % of NT sirna trnsfete ells NT sirna Csp-8 sirna M45 M45mutRHIM
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SUPPLEMENTARY INFORMATION doi:.38/nture59 lmd phosphtse Okdic cid h COS7 SHSY5Y Clf lkline phosphtse Mouse rin c lmd phosphtse Okdic cid h 3P IB () COS7 Supplementry Figure is phosphoprotein., Okdic cid
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Pulldown HeL lyste lyste lyste 25 5 75 5 5kD 37 25 5 MDDIFTQCREGNAVAVRLWLDNTENDLNQGDDHGFSPLHWACREGRSAVVEMLIMRGARINVMNRGDDTP LHLAASHGHRDIVQKLLQYKADINAVNEHGNVPLHYACFWGQDQVAEDLVANGALVSICNKYGEMPVDKA KAPLRELLRERAEKMGQNLNRIPYKDTFWKGTTRTRPRNGTLNKHSGIDFKQLNFLTKLNENHSGELWKG
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doi:10.1038/nture09470 prmt5-1 prmt5-2 Premture Stop Codon () GGA TGA PRMT5 Hypocotyl Length (Reltive to Drk) 0.5 0.3 0.1 ** 30 *** c prmt5-1 d ** *** 150 prmt5-2 ** *** 28 100 26 24 50 prmt5-1 prmt5-2
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BRC repet RPA DSB RAD52 DSB Repir doi:1.138/nture9399 Gp Repir ssdna/dsdna junction ssdna/dsdna junction RPA Binding Resection RPA Binding Filment Formtion or or Filment Formtion DNA Piring DNA Piring
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SUPPLEMENTARY INFORMATION doi:.8/nture99 Supplementry Tle : Primers nd proes. Sequences re written in 5 - direction. Vector construction cirs-7 forwrd cirs-7 reverse cirs-7ir forwrd cirs-7ir reverse cirs-7fs
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nm nm nm nm nm nm. Seed surface. oi-ab. oi-ad. ii-ab. ii-ad/endothelium. endosperm.
B 360-370nm Seed surfce oi- 90-100nm A 630-640nm oi-d ii- ii-d/endothelium 230-240nm 220-230nm 240-280nm 1mm endosperm C oi-d D ii-d/endothelium ii- endosperm Supplementry Figure 1 Cell wll thickness mesurements
Supplementary Materials
Supplementry Mterils Supplementry Figure 1 Supplementry Figure 2 in Fh deficient mice. Supplementry Figure 3 Supplementry Figure 4 Supplementry Figure 5 Supplementry Figure 6 Supplementry Figure 7 Supplementry
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SI Fig. PrpS is single copy gene k 3. 9... EcoRV EcoRV k 5 BmH Pst c well k HindIII HindIII HindIII.3.5 3.. Southern lots of Ppver genomic DNA from plnts with SS8 hplotypes, hyridized with PrpS proe..
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% chnge in ody mss. -.. Smll intestinl mss (g) -1. -. -. Villi length in proximl jejunum (µm) 1 # of crypts/ mm of jejunum g Proximl jejunum # of enterocytes in jejunl villi 1 1 e. -. d Smll intestinl
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DOI: 0.038/n2228 HepG2 (ell pellet ml) Nuler Extrts (.8 mg) -epenent intertnts HepG2 (ell pellet 55.5 ml) Nuler Extrts (227.7 mg) glutthione sephrose F.T. glutthione sephrose F.T. oun F.T. oun F.T. -FXR
Supplementary Fig
Supplementry Fig. 1 * 180-115- 82-64- 49-37- * 180-115- 85-64- 49-37- 26-26- Mem Cyt Mem Cyt Supplementry Fig.1 Specificity of nti-tie2 ntiodies. HUVECs were homogenized nd centrifuged t 400 000g to otin
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a b c Nature Neuroscience: doi: /nn.3632
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British Journal of Pharmacology (2004) 143, & 2004 Nature Publishing Group All rights reserved /04 $
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Relative intensity Relative intensity Relative intensity Supplementary figures a None Caffeine None Caffeine c None Caffeine 6 6 6 ISG 6 6 6 UBE1L 6 6 6 UBCH8 6 6 6 EFP 1 1 DOX (h) 1 1 CPT (h) 1 1 UV (h)
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