Sepantronium bromide ym155

All reagents were purchased from Sigma-Aldrich (St Louis, MO, USA), unless indicated. Primary antibodies for p-mTOR (Ser2448), mTOR, p-S6 (Ser235/236), S6, p-Rb(Ser780), Atg5, LC3, Bcl2l1, Bcl2, Beclin-1 and cleaved-poly (ADP-ribose) polymerase (PARP) were purchased from Cell Signaling Technology (Danvers, MA, USA). Primary antibody of LAMP2 were purchase from Proteintech (Chicago, IL, USA), Primary antibody of SQSTM1/p62 were purchase from Novus Biological (Littleton, CO, USA). Cyclin D1 antibody was purchased from Santa Cruz (Santa Cruz, CA, USA). Sepantronium bromide (YM155) was purchased from Selleck (Houston, TX, USA).

The esophageal cancer cell lines KYSE410 and KYSE150 were obtained from Dr. Yutaka Shimada at the Hyogo College of Medicine [15 (link)]. Mouse embryonic fibroblasts (MEFs) were obtained from Dr. Zhang, Hongbin at Peking Union Medical College. All cell lines were grown in RPMI supplemented with 10% FBS, 100 U/ml penicillin, and 100 μg/ml streptomycin at 37°C in 5% CO₂.
Sepantronium Bromide (YM155) was purchased from Selleck Chemicals (Houston, TX, USA), dissolved in dimethyl sulfoxide (DMSO) to 10 mmol/L stock solutions, and diluted with culture medium. MitoTracker Red CMXRos was purchased from Invitrogen (Life Technologies, USA) and 2, 7-dichlorofluorescein diacetate (H2DCF-DA) and 40, 6-diamidino-2-phenylindole (DAPI) from Sigma-Aldrich (St. Louis, Missouri, USA). Anti-caspase-3 and anti-caspase-9 antibodies were purchased from Enzo (Life Sciences, NY); γH2AX, Survivin, XIAP, mTOR, AKT, Phospho-AKT, ERK, Phospho-ERK, Phospho-S6, 4EBP1, Phospho-4EBP1, LC3, Beclin and Survivin antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA); Apoptosis-inducing factor (AIF) antibody from Santa Cruz Biotechnology, (Santa Cruz, CA, USA); SLC35F2 antibody from Abcam (Cambridge, MA, USA); PAR and PARP antibodies from BD Pharmingen (BD Biosciences, San Jose, CA); and β-actin antibody from Sigma-Aldrich (St. Louis, Missouri, USA).