Anti phospho p70s6

Media for cell culture, anti-hemagglutinin (HA) polyclonal antibody, and LipofectAMINE Plus reagent were ordered from Invitrogen (Carlsbad, CA, USA). Anti-AMPKα (AMPK α-subunit), anti-phosphorylated AMPKα (Thr 172), anti-ACCα, anti-phospho-ACCα, anti-p70S6, and anti-phospho-p70S6 antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Monoclonal anti-HA was from Roche Molecular Biochemicals (Indianapolis, IN, USA). The anti-myc antibody and the polyclonal anti-HA antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-human synphilin-1 polyclonal antibody was generated using the human synphilin-1 fragment (34–500 aa) as the antigen as previously described [3 (link)]. Anti-actin antibody was from Sigma (St. Louis, MO, USA). Compound C (an AMPK inhibitor) [40 (link)] was from Sigma (Burlington, MA, USA).

Lipofectamine 2000 and tissue culture medium were provided from Invitrogen Life Technologies (Carlsbad, CA). Anti-LC3-B antibody was purchased from Abcam (Cambridge, MA) and anti-α-tubulin antibody, Bafilomycin A1 and Rapamycin were from Sigma (St. Louis, MO). Anti-PI3K, anti-phospho-Akt, anti-Akt, anti-phospho-mTOR, anti-mTOR, anti-phospho-P70S6 and horseradish peroxidase-conjugated secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA). Anti-nitrotyrosine antibody was from Millipore (Darmstadt, Germany). Alexa488-conjugated secondary antibodies were obtained from Invitrogen (Carlsbad, CA).