Anti larg

The samples used for western blotting were separated by SDS-PAGE. After electrotransfer, the polyvinylidene fluoride (PVDF) membranes (Merck Millipore, Darmstadt, Germany) were probed with anti-GEF-H1, p(Thr18/Ser19)-MLC, MLC, anti-GST (Cell Signaling Technology, Inc., Danvers, MA, USA), anti-LARG, anti-Ect2, anti-RhoA, anti-β-actin, anti-α-tubulin, or anti-FLAG (Sigma-Aldrich, St. Louis, MO, USA) antibodies, followed by horseradish-peroxidase-conjugated goat anti-rabbit or rabbit anti-mouse antibodies (Zymed Laboratories, Inc., South San Francisco, CA, USA). The blots were developed using an ECL Western Blotting Kit (GE Healthcare, Little Chalfont, UK).

Anti-Atg5 and anti-LARG antibodies were purchased from Sigma-Aldrich (St. Louis, MO). Anti-paxillin, anti-p62, anti-actin, and anti-tubulin antibodies were purchased from BD Biosciences (San Jose, CA), MBL (Aichi, Japan), Millipore (Billerica, MA), and Invitrogen (Carlsbad, CA), respectively. Anti-GEF-H1, anti-RhoB, and anti-RhoC antibodies were from Cell Signaling Technologies (Danvers, MA). Anti-RhoA and anti-Rac1 antibodies were from Cytoskeleton Inc. (Denver, CO). Anti-LC3, anti-phosho FAK and anti-GAPDH were purcharsed from NanoTools (Teningen, Germany), Abcam (Cambridge, UK) and Santa Cruz Biotechnology (Santa Cruz, CA), respectively. Bafilomycin A1 was obtained from Sigma-Aldrich. Rho inhibitor 1 was obtained from Cytoskeleton Inc. and Chemdea LLC. (Ridgewood, NJ), respectively. All other chemicals were from Wako Co. (Osaka, Japan).