Fig. 3.

GPx3 knockdown inhibits OVCAR3 clonogenicity and survival in anchorage independence. A: Cells (100/well) were seeded onto 12-well plates and stained with crystal violet after 10 days of growth. Data expressed as mean ± SEM of 3 independent experiments (n = 4/group; ANOVA, post hoc **p < 0.01). B: GPx3 knockdown by shRNA was verified by semi quantitative real time RT-PCR. C: The effects of GPx3 knockdown on cell viability were assessed under conditions of anchorage independence (a.i.), by culturing cells in 96 well ultra-low attachment (ULA; 1000 cells/well) plates for indicated times, followed by staining with calcein-AM (2 μM), indicating live cells, and Ethidium homodimer-1 (4 μM) indicating dead cells. Representative images are shown. Scale bar = 100 µm. D: Quantification of live/dead staining using ImageJ. Data expressed as mean ± SEM (n = 4–8/group; One-way ANOVA, Tukey's post hoc, **p < 0.01, ****p < 0.0001).