Fig. 3.

Induction of apoptotic cell death by combined treatment with Romidepsin and JQ1. a Increase of subG1 fraction by combination treatment as determined by flow cytometry. b Flow cytometric analysis of UCCs with indicated treatment (Table 1 lower part) after combined staining with PI and Annexin V. Percentages of viable (lower left), early (lower right), or late (upper right) apoptotic and necrotic (upper left) VM-Cub1 cells subsequent to indicated treatments. c Percentage of early apoptotic cells as measured by Annexin V staining for all UCCs and HBLAK control cells are displayed in bar graphs for the respective treatment. d PARP and Caspase-3 cleavage 48 h after treatment assessed by Western blot analysis. e Four UCCs received the combination treatment with or without the Pan-Caspase inhibitor Q-VD-Oph at 30 μmol/l. All cells received the same concentration of DMSO. Cell viability displayed on the ordinate was measured by ATP assay after 48 h. Differences between J + R treatment and J + R treated plus Caspase inhibitor were analyzed using Student’s t test (***p ≤ 0.001, **p ≤ 0.01)