Gelucire

Whole cell lysates were prepared in lysis buffer (20 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM EDTA, 1% NP40 and 10% Glycerol), resolved by SDS-PAGE and blotted with indicated antibodies. The following antibodies were used in this study: Anti-β-catenin, anti-FAK, anti-phospho-FAK (Tyr397), anti-ERK1/2, anti-SRC and anti-Integrin β1 (Cell Signaling Technology, Danvers, MA); anti-GSK3α/β, anti-phospho-GSK3α/β (Tyr279/216), anti-phospho-ERK1/2, anti-β-Actin and anti-SP1 (Santa Cruz Biotechnology, Dallas, TX). SuperSignal West Pico Chemiluminescent Substrate and SuperSignal Western Blot Enhancer (Thermo Fisher Scientific, Waltham, MA) were used to enhance western signal when needed. Vemurafenib, Dabrafenib, GDC-0879 (8 (link)), Erlotinib, SCH772984 (9 (link)), LY3009120 (10 (link)) and ICG-001(11 (link)) were purchased from Selleck Chemicals (Houston, TX). PF-562271 (12 (link)) was purchased from MedKoo Biosciences (Chapel Hill, NC). PLX7904 (13 (link)) was obtained from Plexxikon (Berkeley, CA). Gelucire is a gift from Gattefosse (Paramus, NJ). NE-PER Nuclear and Cytoplasmic Extraction Reagents from Thermo Fisher Scientific were used for subcellular fractionation according to manufacturer’s protocol.