Phospho mtor p mtor

Preparation of cell lysates and details of western blot were described previously^{43 (link)}. The antibodies used included E-cadherin (1:5000; Proteintech, 20874-1-AP), β-Catenin (1:1000, Cell Signaling Technology, CST9562), vimentin (1:1000, Cell Signaling Technology, CST5741), LC3A/B (1:1000, Cell Signaling Technology, CST4108), caspase-3 (1:1000, Cell Signaling Technology, CST9662), cleaved caspase-3 (1:1000, Cell Signaling Technology, CST 9661), PARP (1:1000, Cell Signaling Technology, CST9532), cleaved PARP (1:1000, Cell Signaling Technology, CST 5625), phospho-AKT (p-AKT) (1:2000, Cell Signaling Technology, CST4060), phospho-mTOR (p-mTOR) (1:1000, Cell Signaling Technology, CST5536), mTOR (1:1000, Cell Signaling Technology, CST2972) and AKT (1:2000, Cell Signaling Technology, CST2920). The reaction was visualized using Clarity Western ECL substrate (Bio-Rad, Hercules, CA, USA) and detected by exposure to autoradiographic film.

Cisplatin (DDP, Cat.^#H20010743) injection was purchased from Jiangsu Hausen Pharmaceutical Co., Ltd (Jiangsu, China). Dulbecco's modified Eagle's medium (DMEM, Cat.^#11965092), fetal bovine serum (FBS, Cat.^#10270106), penicillin/streptomycin (Cat.^#10378016), and phosphate-buffered saline (PBS, Cat.^#10010023) were supplied by Gibco (NY, USA). Primary antibodies against Beclin1 (Cat.^#11306-1-AP), p62 (Cat.^#66184-1-Ig), Cytochrome c (cyto. C, Cat.^#66264-1-Ig), and DAPK1 (Cat.^#25136-1-AP) were purchased from Proteintech (Wuhan, China). Primary antibodies against Phospho-AMPKα (p-AMPKα, Cat.^#50081S), Phospho-mTOR (p-mTOR, Cat.^#5536T), Atg7 (Cat.^#8558T), LC3A/B (Cat.^#12741S), Cleaved Caspase-9 (c-Casp 9, Cat.^#20750S), Cleaved Caspase-3 (c-Casp 3, Cat.^#9661T), Cleaved PARP (c-PARP, Cat.^#5625T), Phospho-SAPK/JNK (p-JNK, Cat.^#9255S), Phospho-p53 (p-p53, Cat.^#9284T), Bax (Cat.^#2772T), and GAPDH (Cat.^#5174T) and rabbit (Cat.^#7074P2) or mouse (Cat.^#7076P2) secondary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

FGF21 (P6101) was obtained from Beyotime Biotechnology (Jiangsu, China), chloroquine (CQ) was obtained from Sigma-Aldrich Chemical Company (Milwaukee, WI, USA), and nicotinamide (NAM) was purchased from MedChemExpress (NJ, USA). The primary antibodies directed against Collagen II, MMP-13, p16INK4a, p21WAF1, Lamin B1, and GAPDH were purchased from Abcam (Cambridge, MA, USA), while the primary antibodies directed against Bax, Bcl-2, SQSTM1/p62, and LC3 were obtained from ProteinTech (Wuhan, China). The antibodies against cleaved caspase3 (C-caspase 3), TFEB, mTOR, Phospho-mTOR (p-mTOR), and SIRT1 were procured from Cell Signaling Technologies (Danvers, MA, USA).