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. 2014 Dec 8;9(12):e113479. doi: 10.1371/journal.pone.0113479

Figure 3. U12-induced apoptosis in SMMC-7721.

Figure 3

Morphological and quantitative changes in SMMC-7721 cells after being (A) left untreated, (B) treated with 100 µM U12 for 24 h, or (C) pretreated with 50 µM Z-VAD-fmk or (D) 20 µM Z-IETD-fmk for 1 h. (E) Western blotting was used to estimate PARP cleavage from the 100 µM U12 for 24 h treatments. (F) Detection of apoptotic SMMC-7721 cells in the presence of 80 µM U12 for 2 h and 7 h using Annexin V-FITC/PI analysis. (G&H) Activation of caspase-3, -8, and -9 was evaluated using a caspase activity kit after indicated concentration of U12 treatment at 2 h and 7 h, respectively. (I) Western blot analysis of PARP cleavage on SMMC-7721 cells untreated and treated with indicated concentration of U12 at 12 h.