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Effects of inhibition of caspases by Z-VAD-FMK on apoptosis, expression of Mcl-1 and p21, and phosphorylation of Akt and JNK.

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posted on 20.02.2013, 08:42 authored by Ning Gao, Senping Cheng, Amit Budhraja, E-Hu Liu, Jieping Chen, Deying Chen, Zailin Yang, Jia Luo, Xianglin Shi, Zhuo Zhang

U937 cells were pretreated with the caspase inhibitor Z-VAD-FMK (20 µM) for 1 h, followed by treatment with 80 µM DIM for 12 h and 24 h. (a) Cells were stained with Annexin V/PI, and apoptosis was determined using flow cytometry. **Values for cells treated with DIM and Z-VAD-FMK were significantly reduced compared to values obtained for DIM alone by Student's t-test, p<0.01. (b–d) Total protein extracts were prepared and subjected to Western blot assay using antibodies as indicated.