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. 2023 Feb 3;11:1124960. doi: 10.3389/fcell.2023.1124960

FIGURE 3.

FIGURE 3

OGG1 inhibitors TH5487 and SU0268 increase the effects of the chemotherapeutical drug Etoposide, independently of OGG1. (A) U2OS WT and OGG1 KO cells were exposed to Etoposide alone or in combination with 10 µM of TH5487, SU0268 or Verapamil. Immediately after exposure to the drugs, cells were fixed and DSB were visualized by immunofluorescence using an antibody against γH2AX (green). Nuclei were stained with Hoechst 33342 (blue). Statistical analysis was performed with a Kruskal Wallis test. A segmentation algorithm was used to quantify γH2AX foci number in at least 1000 cells for each condition. Scale bar 50 µm. (B) Distribution of γH2AX foci number per nuclei is presented for a representative experiment out of three. Statistic analysis was performed with a Kruskal Wallis test (***)p < 0.001; (****)p < 0.0001. (C) The number of γH2AX foci per nuclei was normalized to the number of foci in cells exposed to Etoposide and the mean from three independent experiments is represented in the graph. Statistical analysis was performed with a Kruskal Wallis test (**)p < 0.01; (***)p < 0.001; (****)p < 0.0001. (D) Cell survival was evaluated in U2OS WT and OGG1 KO cells, 4 days after exposure to a single treatment with Etoposide or a combined treatment with Etoposide and 10 µM of TH5487, SU0268 or Verapamil. The number of cells were normalized to the untreated control cells. The mean of three independent experiments is represented in the graph. Statistical analysis was performed with a Kruskal Wallis test (*)p < 0.05; (**)p < 0.01; (***)p < 0.001; (****)p < 0.0001.