Fig. 4: A compound screen for elimination of therapy-induced senescent HGSOC cells.

A Schematic representation of primary drug screen design was created with Biorender.com. B Waterfall plot of Z-scores for 1131 compounds evaluated in cisplatin (CP) and CX-5461 (CX) senescent cells. C Dose–response curves for ferroptosis inducers ML-210, RSL3, FIN56 and N6F11. Dashed lines indicate dose–response curves with the inclusion of 1 μM liproxstatin. D Representative fluorescent images of OVCAR8 CP TIS cells exposed to RSL3 in the absence or presence of 1 μM liproxstatin. DAPI staining was used to mark nuclei and rhodamine-phalloidin for cell morphology. Cells were only considered if they were positive for DAPI and phalloidin staining. Scale bar = 50 µm. E Quantification of median fluorescence intensity (MFI) of lipid peroxidation as measured by C11-BODIPY staining is shown as mean ± SEM from n = 3 independent experiments. Statistical analysis was performed using one-way ANOVA with a Šídák’s multiple comparisons test. *p < 0.05; **p < 0.01; ****p < 0.0001; ns not significant. F Quantification of the percentage cell death as measured by fixable NIR staining is shown as mean ± SEM from n = 3 independent experiments. Statistical analysis was performed using one-way ANOVA with a Šídák’s multiple comparisons test. *p < 0.05; ***p < 0.001; ****p < 0.0001; ns not significant. G Representative images of colony formation assay of OVCAR8 cells treated with cisplatin for 48 h, followed by washout. At six days post-washout, cells were treated with control or 100 nM RSL3 for 24 h, then imaged weekly for an additional two weeks.