Rhodamine 123 mitochondrial specific fluorescent dye

Human lung squamous carcinoma cell line SK-MES-1 was purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). Isoalantolactone was purchased from the Chinese materials research center (Beijing, China), dissolved in Dimethyl Sulfoxide (DMSO), which was purchased from Shenggong Company (Shanghai, China). Fetal bovine serum (FBS) was purchased from Gibco (Carlsbad, CA, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT), Hoechst 33342, Dulbecco's Modified Eagle's Medium (DMEM) and Rhodamine 123 mitochondrial specific fluorescent dye were purchased from Sigma (St. Louis, MO, USA). Cell Cycle Analysis Kit (PI+Rnase A), Annexin V-FITC Apoptosis Detection Kit, Reactive Oxygen Species Assay Kit and BCA Protein Assay Kit were purchased from Keygene Company (Nanjing, China). Polyclonal antibodies against β-actin(1:2000 dilution #4967S), Bax(1:1000 dilution #2772S), Bcl-2(1:1000 dilution #2876S), pro-caspase-3(1:1000 dilution #9662P), poly AIsoalantolactone-ribose polymerase (PARP) (1:1000 dilution #9542S), pRb(1:1000 dilution #9306S), p27(1:1000 dilution #2552), p53(1:1000 dilution #9282S), and horseradish peroxidase-conjugated secondary antibodies (goat-anti rabbit, mouse) were purchased from Cell Signaling Technology, Inc. (Shanghai, China). Western Blotting detection kit was purchased from Milipore (Billerica, MA, USA).