Supplementary Table 1. Characterization of HNSCC PDX models established at MSKCC
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1 Supplementary Table 1. Characterization of HNSCC PDX models established at MSKCC Supplementary Table 2. Drug content and loading efficiency estimated with F-NMR and UV- Vis Supplementary Table 3. Complete blood count of mice 24 h following i.v. injection of 25 mg/kg FiBYL719
2 Supplementary Table 4. Antibodies used in western blot. Antibody Supplier Catalog # Phospho-p44/42 MAPK (Erk1/2) (T202/Y204) p-s6 Ribosomal protein (S235/236) p-s6 Ribosomal protein (S240/244) Cleaved PARP (D214) mouse specific Pir (p-igf-irbeta (Y1135/1136) InsR beta 19H7 4370S 2211S 2215S 9544S 3024S Filamin Millipore CBL228 phospho-histone H2A.X (S139), clone JBW301 Millipore β-actin (C4) Santa Cruz SC47778 p44/42 MAPK (Erk1/2) 9102 S6 Ribosomal Protein (5G10) 2217 Phospho-GSK-3β (Ser9) 5558 Rabbit IgG HRP-linked (secondary) Bovine anti-mouse IgG-HRP linked (secondary) Santa Cruz 7074P2 BO415
3 Supplementary Figure 1. P-selectin abundance and p-akt status in pre-clinical HNSCC models. (a) Representative images of immunohistochemistry staining for P-selectin and pakt in different HNSCC cell lines and patient-derived xenografts (PDX), scale bar, 100 µm. (b) Representative images of immunofluorescent staining for CD-31 (red) and P-selectin (green) in H22 PDX. Scale bar, 50 µm.
4 Supplementary Figure 2. Preparation of FiBYL719 nanoparticles using nano precipitation methods.
5 Supplementary Figure 3. Characterization of BYL719-encapsulated nanoparticles. (a) Scanning electron microscopy (SEM) images of P-selectin-targeted FiBYL719 and control DexBYL719 nanoparticles. Scale bar = 100 nm. (b) Nanoparticle diameters measured with dynamic light scattering (DLS). (c) Nanoparticle zeta potential measured with electrophoretic light scattering. (d) Nanoparticles stability over time in growth medium containing 10% fetal bovine serum evaluated by dynamic light scattering (DLS). (e) Drug release profile of BYL719 from FiBYL719 nanoparticles over time at PBS buffers of ph 5.5 and 7.4. (f) Fluorescence images of bovine aortic endothelial cells (BAEC) monolayer treated with either TNFɑ or ionizing radiation (4 Gy) for induction of P-selectin. Near-infrared dye (red) in FiBYL719 and DexBYL719 nanoparticles. CellMask membrane stain (green). DAPI nuclear stain (blue). Scale bar, 15 µm.
6 Supplementary Figure 4. Establishment and characterization of orthotopic HNSCC model. (a) Bioluminescence imaging of mice 7 days following orthotopic tongue engraftment of GFP-Luc- Cal-33 cells using intraperitoneal injection of D-Luciferin (50 mg/ml). (b) In vivo imaging of primary orthotopic tongue GFP-Luc-Cal-33 xenograft using fluorescent stereoscopy. (c) In vivo imaging of cervical lymph node metastasis from orthotopic tongue GFP-Luc-Cal-33 xenograft using fluorescent stereoscopy. (d) H&E and immunohistochemistry staining for Pan-cytokeratin (CK-PAN) in cervical lymph node containing metastatic Cal-33 cells (arrows), scale bar, 1000 μm. (e) Representative images of immunohistochemistry staining for CD-31 and P-selectin in orthotopic Cal-33 tongue xenograft, scale bar, 100 μm.
7 Supplementary Figure 5. Un-cropped western blots from (a) Figure 2, Cal-33 xenografts at different time points following treatment with 25 mg/kg BYL719 or 25 mg/kg FiBYL719, n=3. (b) Figure 3, western blot of γh2ax and cleaved PARP in H22 and H31 patient-derived xenografts 24h post treatment with RT (4 Gy) or RT and 50mg/kg BYL719 or 25 mg/kg FiBYL719 (n=3).
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Supplementary Figure (OH) 22 nanoparticles did not affect cell viability and apoposis. MDA-MB-231, MCF-7, MCF-10A and BT549 cells were
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marker. DAPI labels nuclei. Flies were 20 days old. Scale bar is 5 µm. Ctrl is
Supplementary Figure 1. (a) Nos is detected in glial cells in both control and GFAP R79H transgenic flies (arrows), but not in deletion mutant Nos Δ15 animals. Repo is a glial cell marker. DAPI labels
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DOI:.38/ncb3399 a b c d FSP DAPI 5mm mm 5mm 5mm e Correspond to melanoma in-situ Figure a DCT FSP- f MITF mm mm MlanaA melanoma in-situ DCT 5mm FSP- mm mm mm mm mm g melanoma in-situ MITF MlanaA mm mm
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Figure 1 A 2.0. Akt308 Akt Ctl 1h 3h 6h 9h 12h. β-cat. GSK3β. pakt308. pgsk3β/edu E-cad/pAkt308. Akt1. Actin.
Figure 1 pβcat552 β-cat pgsk3β GSK3β 110 110 pgsk3β/du -cad/ TL 20 µm 20 µm pkt473 pankt pkt473 pankt (days) 0 1 2 3 4 (h) 0 1 3 6 9 12 pβ-cat552/nuclei ensitometricanalysis (Pixel intensity) F ensitometricanalysis
X P. Supplementary Figure 1. Nature Medicine: doi: /nm Nilotinib LSK LT-HSC. Cytoplasm. Cytoplasm. Nucleus. Nucleus
a b c Supplementary Figure 1 c-kit-apc-eflu780 Lin-FITC Flt3-Linc-Kit-APC-eflu780 LSK Sca-1-PE-Cy7 d e f CD48-APC LT-HSC CD150-PerCP-cy5.5 g h i j Cytoplasm RCC1 X Exp 5 mir 126 SPRED1 SPRED1 RAN P SPRED1
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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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T H E J O U R N A L O F C E L L B I O L O G Y
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human epithelial cells were pretreated with control sirna (50 nm) or GSK-3β sirna (50 nm)
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Figures S1-S5, Figure Legends, Table S1 List of primers used in the study
Insulin receptor alternative splicing is regulated by insulin signaling and modulates beta cell survival Pushkar Malakar,4, Lital Chartarifsky,4, Ayat Hija, Gil Leibowitz 3, Benjamin Glaser 3, Yuval Dor,
Nature Neuroscience: doi: /nn Supplementary Figure 1
Supplementary Figure 1 EGFR inhibition activates signaling pathways (a-b) EGFR inhibition activates signaling pathways (a) U251EGFR cells were treated with erlotinib (1µM) for the indicated times followed
Protection against doxorubicin-induced myocardial dysfunction in mice by cardiac-specific expression of carboxyl terminus of hsp70-interacting protein
Protection against doxorubicin-induced myocardial dysfunction in mice by cardiac-specific expression of carboxyl terminus of hsp70-interacting protein Lei Wang 1, Tian-Peng Zhang 1, Yuan Zhang 2, Hai-Lian
ANGPTL2 increases bone metastasis of breast cancer cells through. Tetsuro Masuda, Motoyoshi Endo, Yutaka Yamamoto, Haruki Odagiri, Tsuyoshi
Masuda et al. Supplementary information for ANGPTL2 increases bone metastasis of breast cancer cells through enhancing CXCR4 signaling Tetsuro Masuda, Motoyoshi Endo, Yutaka Yamamoto, Haruki Odagiri, Tsuyoshi
Real-time imaging reveals the single steps of brain metastasis fo mation r
Real-time imaging reveals the single steps of brain metastasis fo mation r Yvonne Kienast, Louisa von Baumgarten, Martin Fuhrmann, Wolfgang E.F. Klinkert, Roland Goldbrunner, Jochen Herms and Frank Winkler
Supplementary Materials for
www.sciencesignaling.org/cgi/content/full/6/278/rs11/dc1 Supplementary Materials for In Vivo Phosphoproteomics Analysis Reveals the Cardiac Targets of β-adrenergic Receptor Signaling Alicia Lundby,* Martin
Figure S1: Effects on haptotaxis are independent of effects on cell velocity A)
Supplemental Figures Figure S1: Effects on haptotaxis are independent of effects on cell velocity A) Velocity of MV D7 fibroblasts expressing different GFP-tagged Ena/VASP family proteins in the haptotaxis