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2 protocols using zm336372

1

Antibody and Drug Reagents for EGFR, AXL, and MAPK Signaling

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Antibodies against EGFR (#2232, RRID:AB_331707), AXL (#8661, RRID:AB_11217435), phospho-ERK (#9101, RRID:AB_331646), ERK (#9102, RRID:AB_330744), PTEN (#9552, RRID:AB_10694066), β-actin (#4967, RRID:AB_330288), GAPDH (#2118, RRID:AB_561053), and vinculin, were obtained from Cell Signaling Technology. Anti-CBL (#05-440, RRID:AB_2290887) was obtained from MilliporeSigma. Anti-NF1 (A300-140A; RRID:AB_2149790) was obtained from Bethyl Antibodies. Vemurafenib, cobimetinib, neratinib, dabrafenib, trametinib, ZM336372, LY3009120, and belvarafenib were obtained from SelleckChem. Drugs for in vitro studies were dissolved in DMSO to create 10 or 100 mmol/L stock solutions and stored at −20°C.
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2

RAS Pathway Inhibitor Effects on bFGF Depletion

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We examined effects of specific inhibitors related to RAS pathways (Favata et al., 1998 (link), Hall-Jackson et al., 1999 (link), Montagut et al., 2008 (link), Powis et al., 1994 (link), Sebolt-Leopold et al., 1999 (link), Vlahos et al., 1994 (link)) on bFGF-depletion assays and in vitro differentiation. PD184532 and wortmannin were purchased from Sigma. U0126 was obtained from Wako. ZM336372, AZ628, and LY294002 were purchased from Selleck. To exclude the effect of cell toxicity, the concentrations of each compound were selected at the range of lower than 80% cell growth inhibition judged by the percentage of nucleus number relative to the vehicle treatment in bFGF-depletion assays (Figures S5B–S5D).
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