Ppp1r15a

Withaferin A (BC grade, >98%) was purchased from Sangon Biotech (Shanghai, China).3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide (MTT), PI dyes, RNase, Penicillin and streptomycin were obtained from Sigma‐Aldrich. Emricasan was purchased from Targetmol. Foetal bovine serum (FBS) and Dulbecco's Modified Eagle Medium (DMEM) were bought from Gibco. The goat‐anti‐rabbit secondary antibody, rabbit monoclonal Bad, GAPDH, phospho‐Ser (112) Bad, Bim, Bak, Bcl‐xL, caspase 3/7/9, cleaved caspase 3/7/8/9, cleaved PARP1, CDK1, phospho‐Thr (161) CDK1, p21, XBP1‐s and ATF3 were obtained from Cell Signaling Technology. Rabbit polyclonal Bax, Bcl‐2, cyclin A/B, p53, HMOX1, DNJB1, ATF6, ATF4, GRP78, CHOP and PPP1R15A were purchased from Proteintech. The details for ID code and dilution of antibodies are listed in the supplemental materials (Table S1).

Appropriate cells were lyzed in RIPA lysis buffer containing a cocktail of protease inhibitors (Roche) and PMSF. Total protein concentration was determined using the bicinchoninic acid (BCA) assay kit (Ding Guo Biotechnology, Shanghai, China). Antibodies against the following proteins were used for immunoblotting: phospho-Akt (Thr308) (#2965S , dilutions 1:1000), phospho-Akt (Ser473) (#4060S, dilutions 1:1000), Akt (#4691L, dilutions 1:1000), phospho-PDK1 (Ser241) (#3438s, dilutions 1:1000), and PDK1 (#3062, dilutions 1:1000), phospho-GSK-3β (Ser9) (#9323P, dilutions 1:1000), GSK-3β (#9315L, dilutions 1:1000), β-catenin (#8480, dilutions 1:1000), phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (#4370S, dilutions 1:1000), p44/42 MAPK (Erk1/2) (#4695S, dilutions 1:1000), cleaved caspase 3 (#9315L, dilutions 1:1000), all from Cell Signaling Technology (MA, USA). PARP1 (#13371-1-AP, dilutions 1:1000), PPP1R15A (#10449-1-AP, dilutions 1:500), (ProteinTech, Rosemont, IL, USA); PPP1α (#271762, dilation 1:1000) and β-actin (#47778, dilutions 1:2000), (Santa Cruz Biotechnology, Santa Cruz, CA, USA); YY1 (#61779, dilutions 1:500, Active Motif, CA, USA). The immunoblots were scanned using an Odyssey infrared imaging system (LI-COR). Immunolabeling was detected using the ECL reagent (Sigma). Protein expression was normalized against β-actin.