Fig. 7

Cell Death induced by ONC212 alone or in conjunction with Navitoclax is augmented by caspase-9 inhibitor. a. Western blot showing the cleaved caspase-9 bands in HeLa and A549 cancer cells treated with ONC212 alone or its combination with Navitoclax. b and c. Percentage of cell viability is shown by MTT assay in HeLa and A549 cancer cells treated with the drugs in the presence and absence of caspase-9 inhibitor Ac-LEHD-CMK (50 μM). Etoposide is used as a positive control drug. (n = 3, mean ± SEM). d, e, f and g. Comparative nuclei condensation based on Hoechst-staining in HeLa and A549 cells treated with ONC212, Navitoclax and a combination of both, in the presence or absence of Ac-LEHD-CMK. The percentage of condensed nuclei in random fields is represented graphically (n = 3, mean ± SEM). h. A schematic diagram describing the functionality of GFP sensor caspase-3-like protease activity indicator (GC3AI) to monitor intracellular caspase-3 activity. Activated caspase-3 cleaves the DEVD linker sequence of the probe which in turn allows masked GFP to fluoresce. i. Flow cytometry scatter plot representing the fluorescence intensity of transiently expressing GC3AI in HeLa cells. Ac-LEHD-CMK enhanced the GFP fluorescence in ONC212 alone or in combination with ONC212 and Navitoclax-treated cells. Etoposide was used as the positive control