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. 2023 Aug 30;12(17):2175. doi: 10.3390/cells12172175

Figure 4.

Figure 4

mH2A1 recruitment to DSBs correlates with chromocenter decondensation. (A) IF confocal images of 0 h-, 16 h-, or 24 h-transfected cells co-expressing Cas9-GFP and MajS-gRNAs, stained with Hoechst and antibodies specific for mH2A1 and γH2AX. Scale bar = 10 μm. (B) Line plot showing the percentage of cells with mH2A1 foci at chromocenters after different time points of Cas9-MajS transfection (0 h, 16 h, 24 h, 48 h, and 96 h). Percentages are based on Cas9-MajS-positive cells. One biological experiment was performed for each time point. (C) Quantifications of the mean fluorescence, area, perimeter, and circularity of chromocenters (Hoechst-dense labeling) after 16 h of transfection without (no Cas9-MajS 16 h) and/or with MajS gRNA (Cas9-MajS 16 h). The number of cells analyzed for each condition is given (n). Each point corresponds to the mean number of foci per cell. Wilcoxon tests were used to assess the significance of the observed differences. * p < 0.05, *** p < 0.001, **** p < 0.0001. (D) IF confocal images of 48 h post-transfected cells co-expressing dCas9-VPR and MajS gRNA or not, stained with Hoechst and antibodies specific for mH2A1 and HP1α. Scale bar = 10 μm.