Figure 6.

The PARP inhibitors PJ34 and MCD113 induced apoptotic tumor cell death in spheroid co-cultures. Co-culture spheroids were generated with JIMT-1-EGFP (green) and Cell Tracker Blue-stained derived MΦs at a 1:1 ratio. Spheroids were kept in cultures for a duration of 4 days (1 day in the presence of 20 ng/mL IFN-γ) and were then treated with 25 μM PJ34 or 55 μM MCD113 for 24 h. Cells were stained with Annexin V 647 to detect apoptosis. Images were taken after spheroid formation on day 5 following PJ34 and MCD113 treatments ((A,B) respectively). Images of 3 spheroids/treatment were taken and analyzed for the fluorescence intensity of Annexin V-Alexa 647 in tumor cells/each well (C,D). Means ± SEM of 3 independent experiments are shown. The statistics were calculated with one-way ANOVA followed by Dunnett’s post-hoc test (* p < 0.05).