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. 2018 Jun 15;9(46):28030–28041. doi: 10.18632/oncotarget.25555

Figure 2. Roniciclib stimulates caspase-3 activity and causes apoptosis in MTC cells.

Figure 2

(A) Caspase-3 activity was evaluated using a fluorometric assay kit in cells treated with roniciclib (100 nmol/L) or vehicle for 24 h in TT and DRO81-1 cells. (B) Cells were treated with roniciclib (100 nmol/L) or placebo for 24 h and stained with fluorescent antibodies against DAPI (blue), cleaved caspase-3 (red) and α-tubulin (green). Cells with cleaved caspase-3 expression are identified (arrowhead). (C) The percentages of cells with cleaved caspase-3 expression were assessed after treatment with placebo or roniciclib (100 nmol/L) for 24 h. Cells were stained with cleaved caspase-3 and its expression was evaluated using immunofluorescence confocal microscopy. A minimum of 1176 cells was counted for each condition. Roniciclib significantly increased the proportion of cells with cleaved caspase-3 expression in TT and DRO81-1 cell lines. (D) Sub-G1 apoptotic cells were detected by measuring the DNA content using flow cytometry in cells treated with roniciclib (100 nmol/L) or vehicle for 24 h and 48 h. Roniciclib increased the proportion of sub-G1 cells in the two MTC cell lines. Scale bar, 20 μm.