Actinomycin d hy 17559

The cell lines, used in this study, were acquired from the Chinese Academy of Sciences (Shanghai, China). The human BCa cell lines namely, RT4, T24, UMUC3, EJ, and SW780 were used. The immortalized human normal bladder epithelial cell line, SV-HUC-1 was also used. Here, the RT4 cell line was cultured in the McCoy's 5A medium (Thermo Fisher Scientific, Inc. USA), SV-HUC-1 in the F12k medium (Sigma-Aldrich, St. Louis, MO, USA), and T24, UMUC3, EJ, and SW780 cell lines were cultured in the RPMI-1640 medium (Thermo Fisher Scientific, Inc. USA). All cell culture mediums were supplemented with 10% fetal calf serum (FCS; Thermo Fisher Scientific, Inc. USA) and 1% penicillin /streptomycin (Hyclone, Logan, UT, USA). These cell lines were incubated in an incubator containing 5% CO₂ at 37°C. Other compounds like chloroquine (CQ, HY-17589A), necrostatin 1 (nec-1, HY-15760), erastin (HY-15763), ferrostatin-1 (fer-1, HY-100579), RSL3 (HY-100218A), NCT-502 (HY-117240), actinomycin D (HY-17559), and MG-132 (HY-13259) were purchased from MedChemExpress (Shanghai, China).

RPMI 1640 medium, fetal bovine serum (FBS), and penicillin-streptomycin were purchased from Gibco (Grand Island, NY). The protein synthesis inhibitor cycloheximide (HY-12320), proteasome inhibitor MG132 (HY-13259), DNA transcription inhibitor actinomycin D (HY-17559), and Bobcat339 (HY-111558A) were obtained from Medchemexpress (Monmouth Junction, NJ). The detailed information about antibodies used in this study was shown in Table S1. PC3, DU145, and LNCaP cells were obtained from ATCC, and were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin.