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. 2022 Jan 29;23(3):1590. doi: 10.3390/ijms23031590

Figure 7.

Figure 7

Role of caspases in MHY2245-induced apoptosis. (A) The expression of pro-caspases-3, -8, and -9 in MHY2245-treated cells was detected via Western blot analysis. β-actin was used as a loading control. Results from three independent experiments are shown. (B) Cell lysates from cells treated with MHY2245 for 24 h were assayed to detect caspase-3, -8, and -9 activities using DEVD-pNA, IETD-pNA, and LEHD-pNA as respective substrates at 37 °C for 1 h. The released fluorescent products were measured. Results are expressed as mean ± SD of three independent experiments. * p < 0.05 and ** p < 0.01 compared with that in vehicle-treated control cells. (C) The presence of cells with sub-G1 DNA content following treatment with MHY2245, indicative of the onset of apoptosis, was detected via flow cytometry. ** p < 0.01 compared with that in vehicle-treated control cells. ## p < 0.01 compared with that in 1.0 μM MHY2245-treated cells. (D) Western blot analysis of PARP and pro-caspase-3 in the total lysates of cells treated with 50 μM Z-VAD-FMK (Z-VAD) and 1.0 μM MHY2245. β-actin was used as a loading control. Representative results from three independent experiments are shown.