3h 2 deoxyglucose

N,N,N’,N’-tetramethyl-p-phenylenediamine dihydrochloride (TMPD), β-actin antibody, oligomycin, rotenone, RU486, ascorbate, succinic acid, digitonin, cyanide 3-chlorophenylhydrazone (CCCP), antimycin A and 4-hydroxytamoxifen (4-HOT) were acquired from Sigma (St Louis, MO, USA). Rapamycin was from AG Scientific, Inc (San Diego, CA, USA); (³H) 2-deoxyglucose was acquired from Amersham Pharmacia Biotech (Piscataway, NJ, USA). Phospho-Akt, Phospho-AMPK, total AMPK, Glut-4, hexokinase I, GPX-1, p70S6K (total and phosphorylated form) and α/β tubulin antibodies were from Cell Signaling Technology, Inc (Danvers, MA, USA). Cytochrome c, Akt total, SCO2, Glut-1, hexokinase II antibodies were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). MitoProfile® Total OXPHOS Human WB Antibody Cocktail was obtained from Mitosciences (Eugene, OR, USA); 7-AAD, MitoSOX™ Red, MitoTracker™ Green and rhodamine-123 were purchased from Invitrogen/Molecular Probes (Carlsbad, CA, USA). p70S6K and MigR1 plasmids were from Addgene (Cambridge, MA, USA) and shRNA P70S6K were from Thermo Scientific/Dharmacon (Lafayette, CO, USA) and were transfected to the cells according to the method provided by the companies.