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. 2024 Jan 18;15(1):69. doi: 10.1038/s41419-023-06418-3

Fig. 4. POLE P286R mutation suppresses the DSB repair and increases cytosolic DNA.

Fig. 4

A Schematic for the fluorescent reporter assay to determine the homologous recombination (HR) pathway. The correction of the DSBs in the target sequence allows GFP expression. B Quantification of the efficiency of HR repair pathway in cells transfected with WT or mutant POLE. C Illustration of fluorescent reporter assay to measure the non-homologous end joining (NHEJ) pathway. D Quantification of the efficiency of NHEJ repair system in cells treated with WT or mutant POLE. E Immunofluorescence analysis of γH2AX level in human EC cells with POLE mutation or WT POLE. Scale bar, 25 μm. F Immunofluorescence staining of dsDNA level in human EC cells with POLE mutation or WT POLE. Scale bar, 25 μm. G Quantification of nuclear γH2AX expression level in the EC cells treated as in (E). H Quantification of dsDNA intensity in the EC cells treated as in (F). I Comet assay of human EC cells with POLE mutations or WT POLE. J Quantification of tail moments in the comet assay. Data are shown as mean ± SD. *p < 0.05, **p < 0.005, ***p < 0.001 significantly different from Control.