Fig. 5

Bcl-2/Bcl-xL inhibitor Navitoclax sensitizes ONC212-mediated apoptosis. a and b. MTT assay showing the comparative cell viability of HeLa and A549 cancer cells treated with ONC212 (24 h) in the presence and absence of Navitoclax (0.25 μM in HeLa and 0.50 μM in A549 for 24 h). c and d. Hoechst-stained fluorescence microscopic images demonstrate comparative nuclei condensation in HeLa and A549 cells induced by ONC212, Navitoclax and a combination of both. Arrows indicate condensed nuclei. The percentage of condensed nuclei in random fields is represented graphically (n = 3, mean ± SEM). e. Representative flow cytometry scatter plots illustrating Annexin V-FITC/PI staining in HeLa and A549 cell lines under various treatment conditions: control, ONC212 (24 h), ONC212 (48 h), and ONC212 + NAV (24 h). The combination of ONC212 with Navitoclax shows a substantial increase in the Annexin V-FITC positive population compared to ONC212 alone at 24 h or 48 h, indicating enhanced apoptosis. f and g. Western blot showing the cleavage of caspase-3 and PARP in HeLa and A549 cells on treatment with ONC212 in the presence and absence of Navitoclax. Etoposide is used as a positive control. Co-treatment with ONC212 and Navitoclax for 24 h synergistically enhanced caspase-3 and PARP cleavage compared to ONC212 treatment for 24 h and 48 h