Fig. 8: Loss of BRCA1, DNA2, MRE11, WRN, but not CtIP, induces the formation of 53BP1 nuclear bodies or micronuclei (MN). | Nature Communications

Fig. 8: Loss of BRCA1, DNA2, MRE11, WRN, but not CtIP, induces the formation of 53BP1 nuclear bodies or micronuclei (MN).

From: DNA double-strand break end resection factors and WRN facilitate mitotic DNA synthesis in human cells

Fig. 8

a Experimental workflow for analyzing 53BP1 bodies or MN in the next G1 phase cells following APH treatment in S-phase and depletion of BRCA1, DNA2, CtIP, MRE11, or WRN by siRNAs in U2OS cells. b Western blot analysis of the expression of BRCA1, DNA2, CtIP, MRE11, or WRN at the end of G2 phase. Tubulin was used as loading control in all cases. This experiment was repeated independently with similar results at least three times. Representative immunofluorescence images (c) and quantification of 53BP1 bodies (d) or MN (e). DNA was stained with DAPI (blue). 53BP1 was detected with antibody and is shown red. MN was shown by DAPI and indicated by white arrows. Scale bars, 10 μm. f Experimental workflow for analyzing 53BP1 bodies or MN in the next G1 phase cells following APH treatment in S-phase and inhibition of the function of MRE11 or DNA2 by PMF39 or C5 respectively. Representative immunofluorescence images (g) and quantification of 53BP1 bodies (h) or MN (i). DNA was stained with DAPI (blue). 53BP1 was detected with antibody and is shown red. MN was shown by DAPI and indicated by white arrows. Scale bars, 10 μm. In each quantification, data are presented as mean ± s.e.m. of at least 3 independent experiments. n = 150: 50 pair of new G1 cells were analyzed in each condition in each replicate. In total, 150 pair of new G1 cells were analyzed for each condition. Statistical p values were calculated with two-tailed non-parametric Mann–Whitney tests and indicated in quantification graphs. Source data are provided as a Source Data file.

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