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. 2021 Mar 25;13(7):1519. doi: 10.3390/cancers13071519

Figure 5.

Figure 5

OTX015 exposure radiosensitizes OC cells by inducing apoptosis and inhibiting colony-forming ability and DNA damage repair. SKOV3 cells, treated with or without OTX015 (1 μM), were exposed or not to radiation (4 Gy). (a) Four h after IR, SKOV3 cells were plated at low concentration and cultured for 12 days. Representative images of colonies marked with crystal violet. Colony formation efficiency was obtained by crystal violet optical density from different assays, each in triplicate. Histograms are means ± SD. Statistical significances were calculated by two-way ANOVA: ** p < 0.01 and *** p < 0.001, vs. DMSO without IR; $$ p < 0.01 vs. DMSO/IR; ## p < 0.01 vs. OTX015 without IR. (b) WB of selected markers of cell cycle arrest (p21) and DNA damage/response (γH2AX, DNA-PK, ATM) in SKOV3 cells with or without OTX015 and/or IR. Tubulin served as normalizer and representative blot was shown. (c) WB of specific proteins involved in apoptosis (PARP, Caspase-3) were performed on SKOV3 cells treated with or without OTX015 and/or IR. Tubulin served as loading control and representative blot was shown. (d) q-PCR analysis (left panel) of GNL3 transcript levels in OTX015/IR-exposed SKOV3 cells, expressed as fold change over control cells (DMSO), set at 1. β-actin mRNA was used as endogenous control. Histograms indicate mean values ± SD of three independent experiments, each in triplicate. Statistical significances were calculated by two-way ANOVA (* p < 0.05 vs. DMSO; $$ p < 0.01 vs. IR). WB analysis (right panel) performed on protein extracts from OTX015/IR-exposed SKOV3 cells. Tubulin served as normalizer. The original Western Blot images can be found in Figure S4.