Akt 4

DMEM/F12 (F12), Trypsin-EDTA, Igepal CA-930, Akt-IV, and SB216763 were purchased from Sigma–Aldrich, Inc. (St. Louis, MO, USA). Fetal calf serum (FCS) was acquired from Equitech-Bio, Inc. (Kerrville, TX, USA). A cocktail of sodium penicillin G, streptomycin sulfate, and amphotericin B was purchased from Gibco-BRL (Gaithersburg, MD, USA). Halt-TM Phosphatase inhibitor cocktail was purchased from Thermo Fisher Scientific (Waltham, MA, USA). Protease inhibitor cocktail was acquired from GE Healthcare Biosciences (Little Chalfont, UK). Trizol reagent and EXPRESS One-Step SYBR GreenER Universal Kit were purchased from Invitrogen (Carlsbad, CA, USA). Bovine IL-10 and IL-8 TSZ ELISA kit were purchased from Biotang (Waltham, MA, USA). Plasmid pLKO.1-GSK3β-#1 was a gift from Alex Toker (Addgene plasmid #32497) and PLKO.1 plasmid was a gift from Bob Weinberg (Addgene plasmid # 8453). pCF CREB M1 was a gift from Marc Montminy (Addgene plasmid # 22969). The generation of pCF empty vector was done by the enzyme digestion of pCF CREB M1 with SacI restriction enzyme, and then the purification and ligation of the 5-kb fragment were generated.

All media and supplements used for cell cultures were purchased from Gibco (Carlsbad, CA, United States), unless otherwise noted. ATP assay kit was obtained from Beyotime (Beyotime, China). LY294002 and Akt-iv was obtained from Sigma-Aldrich (St. Louis, MO, United States). PGC1α siRNA, scrambled siRNA and transfection reagent were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States). RIPA lysis buffer, phenylmethanesulfonyl fluoride (PMSF), and halt phosphatase inhibitor cocktail were purchased from Pierce Biotechnology (Rockford, IL, United States). Antibodies against phospho-Ser473 Akt, phospho-Ser9 GSK3β, Bcl-2 and Bax were obtained from Cell Signaling Technology (Beverly, MA, United States). Antibodies against β-actin, PGC1α, Nrf2 and HO-1 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, United States). All other reagents were from Sigma-Aldrich (St. Louis, MO, United States) except where stated otherwise.