Z devd fmk

The reagents and solvents were purchased commercially and used without further purification unless otherwise noted. 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), Hoechst 33258, 2-(6-Amino-3-imino-3H-xanthan-9-yl) benzoic acid methyl ester, RIPA Lysis Buffer and Annexin V-FITC/PI Apoptosis Detection Kit were purchased from Sigma. The Caspase Activity Assay Kit, caspase-3 inhibitor Z-DEVD-FMK, caspase-9 inhibitor Z-LEHD-FMK and caspase inhibitor Z-VAD-FMK, LDH Release Assay Kit were purchased from Beyotime.

Phosphate-buffered saline (PBS), fetal bovine serum (FBS), RPMI-1640, and trypsin were obtained from Gibco (Grand Island, NY, USA). The A549 cell line was obtained from the American Type Culture Collection (Manassas, VA, USA) and was cultured in RPMI-1640 medium containing 10% FBS and 1% penicillin-streptomycin at 37 °C in an atmosphere of 5% CO₂. Cytochrome c, Bax, Bcl-2, PARP, cleaved PARP, caspase-3, cleaved caspase-3, caspase-9, cleaved caspase-9, p53, p21, Cyclin A2, Cyclin E1, Cyclin B1, CDK2, Erk, p-Erk, JNK, p-JNK, p38, p-p38, mTOR, p-mTOR, Akt, p-Akt, FAK, p-FAK, PI3K, p-PI3K, p62, LC3A/B- I/II, β-actin, GAPDH and all secondary antibodies were acquired from Cell Signaling Technology (Beverly, MA, USA). Necrostatin-1 (Nec-1, an inhibitor of RIP1K), Z-VAD-FMK (an inhibitor of pan caspase), Z-DEVD-FMK (an inhibitor of caspase-3), N-Acetyl-L-cysteine (NAC), mitochondrial membrane potential assay kit for JC-1, reactive oxygen species (ROS) assay kit, cell cycle and apoptosis analysis kit, and DAPI were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Smp24 and FITC-labeled Smp24 were synthesized as reported in our previous study [11 (link)].