Anti pi3 kinase

Nectin-4-MMAE is a gift from Mabwell Bioscience CO., LTD. Shanghai China. Autophagy modulators: LY294002 (Medchemexpress, HY-10108, Shanghai China); Chloroquine (Sigma-Aldrich, C6628, Darmstadt Germany); Rapamycin (Sangon Biotech, A606203, Shanghai China). Cell viability assay: MTT (Beyotime, C0009S, Shanghai, China). ADC labeling: AlexaFlour 488 protein label kit (ThermoFisher, A10235, MA US). Cell apoptosis assay: cell apoptosis detection kit (Meilunbio MA0220-1, Dalian China). Autophagy assay: Cyto-ID autophagy detection kit (Enzo Life Sciences, ENZO-51031-K200, NY US); Lyso-Tracker-DND 99 (Invitrogen, L7528, CA US). Antibodies: Primary antibody: anti-GAPDH, #2118; anti-LC3 I/II, #3868; anti-SQSTM1, #8025; anti-caspase 3, #9662; anti-PARP, #9542; anti-phospho-mTOR (Ser2448), #2971; anti-mTOR, #2983; anti-phospho-AKT (Ser473), #4060; anti-phospho-p70s6 Kinase (Ser371), #9208; anti-p70s6 Kinase, #2708; anti-phospho-4EBP1 (Thr45), #2971; anti-4EBP1, #9644, anti-PI3 Kinase, #4257; anti-phospho-PI3 Kinase, #4228 (Cell Signaling Technology, MA US). Anti-PDK-1, AF7707 (Beyotime Biotechnology, Shanghai, China). Anti-PTEN, A19104 (Abclonal, Wuhan China). Secondary antibody: HRP-conjugated anti-rabbit secondary antibody, #7074; anti-mouse IgG secondary antibody, #7076 (Cell Signaling Technology, MA US).

Fetal bovine serum (FBS), Dulbecco’s modified Eagle’s medium (DMEM) were purchased from Gibco (Gibco, Thermo Fisher Scientific, Waltham, MA, United States). Penicillin and streptomycin were obtained from Beyotime (Beyotime, Sichuan, China). Paclitaxel (PTX) purchased from Sigma-Aldrich (St. Louis, MO, United States) Primary antibodies, including anti-EGFR (#4267), anti-p-EGFR (#3777), anti-PI3-kinase (#4249), anti-p-PI3K (#4228), anti-AKT (#4691), anti-p-AKT (#4060), anti-NFκB-P65 (#8242), anti-p-NFκB-P65 (#3033), anti-MAPK (#4695), anti-p-MAPK (#4370), anti-Cleaved Caspase-3 (#9664), anti-PARP (#9542) and β-actin (#4970) were purchased from Cell Signaling Technology Inc. (CST, MA, United States).