Figure 1 A 2.0. Akt308 Akt Ctl 1h 3h 6h 9h 12h. β-cat. GSK3β. pakt308. pgsk3β/edu E-cad/pAkt308. Akt1. Actin.
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1 Figure 1 pβcat552 β-cat pgsk3β GSK3β pgsk3β/du -cad/ TL 20 µm 20 µm pkt473 pankt pkt473 pankt (days) (h) pβ-cat552/nuclei ensitometricanalysis (Pixel intensity) F ensitometricanalysis (Pixel intensity) µm kt308 kt473 1d 2d 3d 4d kt308 kt473 1h 3h 6h 9h 12h
2 Figure 2 pkt 4 pkt 246 (days) pkt 4 pkt 246 (h)
3 Figure 3 ppk1 PK1 ensitometricanalysis (Pixel intensity) ppk η θ β ε ζ ppk1 ppk-1 PK-1 ontrol θ eta zeta η β ε ζ ensitom: OT- Flag 20 µm
4 Figure 4 ensitometricanalysis (Pixel intensity/) ppk1 PK1 J TG5 Raptor Oct yt In vivo / TNFα Rapt SW480 In vitro +Q yt Q S58/Rapt S58 yt MG132 yt LLN L3 I II TG5 G FLG L3/Nuc K F WT ps58 pan (days) 0 4 IP IgG IP: Flag Input IFN/TNF Raptor ensi: L3 S58 1 (days) H S η ppk1 PK ζ Flag Pon Red S58 WT+ 10 µm I pkt µm Vector 10 µm S58 p62 S58
5 Figure 5 ensitometricanalysis (Pixel intensity) MSO V02 PRP asp-3 L asp-3 H p-hist3 PN 130 /V02 1cm V02 V02/ SW480 PRP asp-3 p-hist3 130 Hist3 15 SW480 V02 V02/ pkt473 pkt473 MSO V02
6 Figure 6 I (isease ctivity Index) asp-3/nuclei TL kt VIII 0.0 (ays): TL /MSO /kti VIII ktinviii 1cm Length (cm) pkt 308 Red Ponc µm /kti VIII asp-3 positive cells/100 crypts pkt473 PRP asp-3 S/kti VIII /kti VIII 130 pβ-cat552 PRP H PRPc L S F In vivo Nucleus ytosol pkt473 /V02 /V02 Hist3 GPH 17
7 Figure 7 SW480 +TNFα Vector η +TNFα +TNFα /GSK asp-3 PRP ppk1 PK η /TNFα Receptor? PI3 K 130 %Piknotic Nuclei SW480 +TNFα PIP3 NS GSK Oct- leaved asp-3 Membrane SW480 +TNFα GSK µm KT1 pkt1308 ps58 PK1 TOR1 Nucleus utophagosome
8 Supplementary figure ay 1 ay 2 ay 3 ay 4 ensitometricanalysis (Pixel intensity) (ays): I (isease ctivity Index) 1 cm induced inflammation decreases cell proliferation and colon length. () ffects of -induced inflammation on cell proliferation markers (PN, phistone3) and disease activity (isease ctivity Index (I)). 57L/6J were treated with 3% dissolved in drinking water for 1-4 days. Relative values for PN and phistone3 obtained from densitometric analysis were normalized to actin. I was analyzed as previously reported (Laukoetteret al, 2007). n=6. () Representative image of colon tissue obtained from control and colitic mice. 57L/6J mice were treated with 3% dissolved in drinking water for 1-4 days. n=6. ar= 1cm.
9 Supplementary figure 2 L3 I II /TNFα 3h 6h pβ-cat552 β-catenin /TNFα 0 3h 6h /TNFα WT S58 S58 5µm pkt473 ps58 F : ps6 G In vivo Rapam ktinhviii W: IP, W: Flag H (4 ) -GFP Input IgG WT-Flag S58-Flag S58-Flag η ppk1 PK ζ Flag Ponceau ps58 10µm S58 and ps58 increased in the mucosa of colitic mice and regulate kt activation. (, upper panel) L3I and L3II were analyzed in SW480 cells treated with cytokines. was used as loading control. (, lower panel) cridine orange (green) was used to analyze the presence of autophagosomes. Nuclei blue. ar= 5µm. () ps58 and p were analyzed in colonic cell lysates of 57L/6J mice injected intraperitoneally with /TNFα. was used as loading control. () WT-Flag, ζWT-Flag and WT-Flag were precipitated from cells cotransfected with a plasmid expressing ζ-GFP plus WT-Flag, WT-Flag or WT-Flag. Westernblot against pan and Flag were performed. (), ppk1, PK1, and Flagwere analyzed in SW480 cells expressing increasing concentrations of ζ S58. Ponceau read was used as loading control. () pβ-cat552, β- catenin,, pkt473, and were analyzedin SW480 cells transfected with ζwt, ζs58 and ζ S58 and treated with (12h). was used as loading control. (F) and ps6 were analyzed in SW480 cellstreated with, plus rapamycin (200 nm) and plus kt InhibVIII (2.12µM). was carried out for 12h. was used as loading control. (G) ζ, p ζ S58 and σ were analyzed in the mucosa of mice treated with 1-4 days. ensitometric analysis is shown. n=6. (H) The distribution of and ps58 was analyzed in the mucosa of coliticmice by immunofluorescence. and ps58, green. Highly enriched cells are marked by arrow () and arrowhead (ps58). iscontinuous line denotes crypt shape. Nuclei, blue. du= red ar= 10µm.
10 Supplementary figure 3 asp-3 L asp-3 H PN TUNL/Nuclei 30µm ensitometricanalysis (Pixel intensity/) PRP cleav asp-3 cleav phist3 (ays): Ponceau M30/Nuclei 24h 48h ps58 3% MSO V02 Inflammation enhances apoptosis in Is. () aspase-3, PN and phist3 were analyzed in control and treated mice. 57L/6J were treated with 3% dissolved in drinking water for 4 days. Low xposure (L) and High xposure (H) for caspase-3 are shown. Ponceau red was used as loading control. () poptosis was analyzed in the mucosa of control and treated mice by Tunelstaining (green). 57L/6J were treated with 3% dissolved in drinking water for 1-4 days. Nuclei, blue. ar=30µm. () Graph shows relative densitometric values for PRP and asp3 cleavage in the mucosa of treated animals. 57L6/J mice received 3% in drinking water for 1-5 days. () poptosis was analyzed in SW480 cells treated with 100U/ml of by M30 staining (green). Nuclei, blue. ar=20µm.() and ps58 levels were analyzed in the mucosa of mice treated with /MSO or /V02.57L6/J mice received 3% in drinking water for 4 days. Mice were injected daily with 10 mg/kg of weight of V02 via peritoneum. was used as loading control.
11 Supplementary figure 4 pkt (Thr308) Surface ottom In vivo Nucleus pkt473/du pkt473 Hist3 pkt473/du PN/Nuclei ktinviii 20µm PRP PN positive cells per crypt Vector Vector myr 130 ktaccumulates in the nucleus of Is and regulates cell proliferation during inflammation. () pkt 473( White) was analyzed by immunofluorescence in samples of mice exposed to 4 days. pkt473 is observed in cytosol in proliferating cells green star and nuclear staining is observed in non-proliferating cells green arrow. du, red. Nuclei, lue. ar= 10µm. () pkt 308 (Green) was analyzed by immunofluorescence in samples of mice exposed to 2 and 4 days. is observed at membrane (White arrow) and nucleus. Nuclei, lue. ar=10µm. () pkt 473, and presence were analyzed in nuclear fractions of Is obtained from mice exposed to 4 days. pkt473, and were enriched in nuclear fractions of colitic mice. Histone 3 was used as marker for nuclear fraction. ()ell proliferation was analyzed in the mucosa of control and mice treated with /MSO or /kt inhibitor VIII by PN staining (red). rypts are marked by dotted line. 57L6/J mice received 3% in drinking water for 4 days. Mice were injected daily with 10 mg/kg of weight of kt inhibitor VII via peritoneum. Nuclei, blue. ar=20µm. Quantification is showed in the graph. p<0.05; p<0.001 (), and PRP cleavage were analyzed in SW480 cells expressing myristoylated(.myr) that were exposed to for 36h. was used as loading control.
12
Supplementary Figure 1.TRIM33 binds β-catenin in the nucleus. a & b, Co-IP of endogenous TRIM33 with β-catenin in HT-29 cells (a) and HEK 293T cells
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Supplementary Figure 1
Supplementary Figure 1 6 HE-50 HE-116 E-1 HE-108 Supplementary Figure 1. Targeted drug response curves of endometrial cancer cells. Endometrial cancer cell lines were incubated with serial dilutions of
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TABLE OF CONTENTS SUPPLEMENTARY LEGENDS... 2 11 MOVIE S1... 2 FIGURE S1 LEGEND... 3 FIGURE S2 LEGEND... 4 FIGURE S3 LEGEND... 5 FIGURE S4 LEGEND... 6 FIGURE S5 LEGEND... 7 FIGURE S6 LEGEND... 8 FIGURE
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a 1% 5% % 1% 5% % OVAb βgal activity A.U. (x1 4 ) 2 1 5 βgal activity A.U. (x1 4 ) 2 1 BSAb 2 hours 4 hours OVAb BSAb OVAb BSAb,1 1 1 1 SIINFEKL (ng/ml) CFSE b Beads Alexa488 (%) 8 6 4 2 ** ** 1:1 5:1
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a CD11c Na + K + ATPase Na + K + ATPase CD11c x-y CD11c Na + K + ATPase Na + K + ATPase CD11c x-z c b x-y view BoNT NAPs CD11c BoNT CD11c NAPs BoNT NAPs CD11c 90 x-z view Apical Basolateral Supplementary
Supplementary Figure 1
Supplementary Figure 1 ZV 50 nm Relative to Protein Levels () C Relative to Protein Levels () 6 4 2 0 10 8 6 4 2 0 Treatment Time (6 h) ZV Concentration (25 µm) ZV Concentration (25 µm) Supplementary Figure
Supporting Information
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Supplementary Fig. S1. Schematic diagram of minigenome segments.
open reading frame 1565 (segment 5) 47 (-) 3 5 (+) 76 101 125 149 173 197 221 246 287 open reading frame 890 (segment 8) 60 (-) 3 5 (+) 172 Supplementary Fig. S1. Schematic diagram of minigenome segments.
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293T cells were transfected with indicated expression vectors and the whole-cell extracts were subjected
SUPPLEMENTARY INFORMATION Supplementary Figure 1. Formation of a complex between Slo1 and CRL4A CRBN E3 ligase. (a) HEK 293T cells were transfected with indicated expression vectors and the whole-cell
Supplementary Materials
Supplementary Materials Supplementary Figure S1 Regulation of Ubl4A stability by its assembly partner A, The translation rate of Ubl4A is not affected in the absence of Bag6. Control, Bag6 and Ubl4A CRISPR
LPS LPS P6 - + Supplementary Fig. 1.
P6 LPS - - - + + + - LPS + + - - P6 + Supplementary Fig. 1. Pharmacological inhibition of the JAK/STAT blocks LPS-induced HMGB1 nuclear translocation. RAW 267.4 cells were stimulated with LPS in the absence
SUPPLEMENTARY INFORMATION
DOI: 10.1038/ncb2607 Figure S1 Elf5 loss promotes EMT in mammary epithelium while Elf5 overexpression inhibits TGFβ induced EMT. (a, c) Different confocal slices through the Z stack image. (b, d) 3D rendering
a) Primary cultures derived from the pancreas of an 11-week-old Pdx1-Cre; K-MADM-p53
1 2 3 4 5 6 7 8 9 10 Supplementary Figure 1. Induction of p53 LOH by MADM. a) Primary cultures derived from the pancreas of an 11-week-old Pdx1-Cre; K-MADM-p53 mouse revealed increased p53 KO/KO (green,
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 Edens and Levy, http://www.jcb.org/cgi/content/full/jcb.201406004/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. Nuclear shrinking does not depend on the cytoskeleton
Supplementary Figures
Supplementary Figures Supplementary Figure 1 Characterization of stable expression of GlucB and sshbira in the CT26 cell line (a) Live cell imaging of stable CT26 cells expressing green fluorescent protein
Supplementary Figure 1: si-craf but not si-braf sensitizes tumor cells to radiation.
Supplementary Figure 1: si-craf but not si-braf sensitizes tumor cells to radiation. (a) Embryonic fibroblasts isolated from wildtype (WT), BRAF -/-, or CRAF -/- mice were irradiated (6 Gy) and DNA damage
(A) PCR primers (arrows) designed to distinguish wild type (P1+P2), targeted (P1+P2) and excised (P1+P3)14-
1 Supplemental Figure Legends Figure S1. Mammary tumors of ErbB2 KI mice with 14-3-3σ ablation have elevated ErbB2 transcript levels and cell proliferation (A) PCR primers (arrows) designed to distinguish
Supplementary Figure 1. The CagA-dependent wound healing or transwell migration of gastric cancer cell. AGS cells transfected with vector control or
Supplementary Figure 1. The CagA-dependent wound healing or transwell migration of gastric cancer cell. AGS cells transfected with vector control or 3xflag-CagA expression vector were wounded using a pipette
Supplementary Figure 1. Validation of astrocytes. Primary astrocytes were
Supplementary Figure 1. Validation of astrocytes. Primary astrocytes were separated from the glial cultures using a mild trypsinization protocol. Anti-glial fibrillary acidic protein (GFAP) immunofluorescent
Prolonged mitotic arrest induces a caspase-dependent DNA damage
SUPPLEMENTARY INFORMATION Prolonged mitotic arrest induces a caspase-dependent DNA damage response at telomeres that determines cell survival Karolina O. Hain, Didier J. Colin, Shubhra Rastogi, Lindsey
Supplementary Materials for
www.sciencesignaling.org/cgi/content/full/4/199/ra75/dc1 Supplementary Materials for Signaling by the Matrix Proteoglycan Decorin Controls Inflammation and Cancer Through PDCD4 and MicroRNA-21 Rosetta
Time after injection (hours) ns ns
Platelet life span (% iotinylated platelets) 1 8 6 4 2 4 24 48 72 96 Time after injection (hours) 6 4 2 IgG GPIα GPIβ GPII GPVI Receptor expression (GeoMean, fluorescence inteity) Supplementary figure
* * A3027. A4623 e A3507 A3507 A3507
a c L A327 d e A37 A37 A37 Supplementary Figure 1. Clinical manifestations of individuals with mutations. (a) Renal ultrasound of right kidney in A327 reveals small renal cysts, loss of corticomedullary
Supplementary Figure 1
Supplementary Figure 1 14 12 SEM4C PLXN2 8 SEM4C C 3 Cancer Cell Non Cancer Cell Expression 1 8 6 6 4 log2 ratio Expression 2 1 4 2 2 p value.1 D Supplementary Figure 1. Expression of Sema4C and Plexin2
Supplementary Materials for
www.sciencesignaling.org/cgi/content/full/2/1/ra81/dc1 Supplementary Materials for Delivery of MicroRNA-126 by Apoptotic Bodies Induces CXCL12- Dependent Vascular Protection Alma Zernecke,* Kiril Bidzhekov,
Supplementary Figure S1. Venn diagram analysis of mrna microarray data and mirna target analysis. (a) Western blot analysis of T lymphoblasts (CLS)
Supplementary Figure S1. Venn diagram analysis of mrna microarray data and mirna target analysis. (a) Western blot analysis of T lymphoblasts (CLS) and their exosomes (EXO) in resting (REST) and activated
effects on organ development. a-f, Eye and wing discs with clones of ε j2b10 show no
Supplementary Figure 1. Loss of function clones of 14-3-3 or 14-3-3 show no significant effects on organ development. a-f, Eye and wing discs with clones of 14-3-3ε j2b10 show no obvious defects in Elav
A. Generation and characterization of Ras-expressing autophagycompetent
Supplemental Material Supplemental Figure Legends Fig. S1 A. Generation and characterization of Ras-expressing autophagycompetent and -deficient cell lines. HA-tagged H-ras V12 was stably expressed in
Irf1 fold changes (D) 24h 48h. p-p65. t-p65. p-irf3. t-irf3. β-actin SKO TKO 100% 80% 60% 40% 20%
Irf7 Fold changes 3 1 Irf1 fold changes 3 1 8h h 8h 8h h 8h p-p6 p-p6 t-p6 p-irf3 β-actin p-irf3 t-irf3 β-actin TKO TKO STKO (E) (F) TKO TKO % of p6 nuclear translocation % % 1% 1% % % p6 TKO % of IRF3
Supplementary Figure 1
Supplementary Figure 1 AAV-GFP injection in the MEC of the mouse brain C57Bl/6 mice at 4 months of age were injected with AAV-GFP into the MEC and sacrificed at 7 days post injection (dpi). (a) Brains
Supplementary Fig. 1. GPRC5A post-transcriptionally down-regulates EGFR expression. (a) Plot of the changes in steady state mrna levels versus
Supplementary Fig. 1. GPRC5A post-transcriptionally down-regulates EGFR expression. (a) Plot of the changes in steady state mrna levels versus changes in corresponding proteins between wild type and Gprc5a-/-