S6k1 inhibitor pf 4708671

Bortezomib and mitomycin C were obtained from Santa Cruz (Dallas, TX, USA). Akt inhibitor (LY294002), S6K1 inhibitor (PF-4708671), and protease inhibitor cocktail were purchased from Sigma Aldrich (St. Louis, MO, USA). Flag-Beclin-1 constructs (wild-type (WT) and S234A/S295A (AA)) were provided by Dr. Beth Levine (University of Texas Southwestern Medical Center, Dallas, TX, USA). Anti-hemagglutinin (HA), anti-caspase 8, anti-caspase 9, anti-caspase 3, anti-phosphorylated Akt (S473)/Akt, anti-autophagy-related protein (ATG)7, anti-BiP, anti-activating transcription factor 4 (ATF4), anti-CCAAT-enhancer-binding protein homologous protein (CHOP), and anti-poly (ADP-ribose) polymerase (PARP) antibody were purchased from Cell Signaling (Danvers, MA, USA). Anti-microtubule-associated protein 1A/1B-light chain 3 (LC3) and anti-actin antibody were purchased from Sigma Aldrich. Anti-Beclin-1 and anti-cytochrome c antibody were purchased from BD PharMingen (San Jose, CA, USA). Phospho-Beclin-1 S234 and S295 antibodies were purchased from PhosphoSolutions (Aurora, CO, USA). Mouse monoclonal anti-ATE1, anti-green fluorescent protein (GFP), and anti-p62 came from Santa Cruz.