Fig. 5: Methyl-USP11 regulates end-resection, ATR signalling and G2/M checkpoint. | Nature Communications

Fig. 5: Methyl-USP11 regulates end-resection, ATR signalling and G2/M checkpoint.

From: Arginine methylation and ubiquitylation crosstalk controls DNA end-resection and homologous recombination repair

Fig. 5

a USP11 and MRE11 are epistatic in the regulation of RPA foci formation after IR (3 Gy) in late S/G2 HeLa cells. CENPF-positive cells were scored for RPA foci formation (mean ± SD; n = 3 independent biological repeats; ****p < 0.0001; one-way ANOVA and Tukey post hoc test). b USP11 is not epistatic with CtIP in the regulation of RPA foci formation after IR (3 Gy) in late S/G2 HeLa cells (mean ± SD; n = 3 independent biological repeats; Student’s t test (two-sided, equal variance): ***p = 0.0096). c Methylated USP11 is epistatic with MRE11 in the regulation of RPA foci formation after IR in late S/G2 MCF7 cells. CENPF-positive cells were scored for RPA foci formation (mean ± SD; n = 3 independent biological repeats; one-way ANOVA and Tukey post hoc test: groups A and B *p = 0.031; groups A–C **p = 0.0046; groups A–D **p = 0.0022; groups A–E *p = 0.0162; groups A–F ***p = 0.0006). d Methylated USP11 is epistatic with MRE11 in the regulation of RPA foci formation after IR in late S/G2 HeLa cells. CENPF-positive cells were scored for RPA foci formation (mean ± SD; n = 3 independent biological repeats; one-way ANOVA and Tukey post hoc test: ****p < 0.0001). e HeLa-USP11-R433K cells display defective ATR signalling. HeLa-USP11 or HeLa-USP11-R433K cells were transfected with siUSP11, exposed to 10 Gy IR and then harvested at the time-points indicated for immunoblot analysis. Representative image of n = 3 independent biological repeats. f HeLa-USP11-R433K cells exhibit a G2/M checkpoint deficiency. Cells were treated with 5 Gy IR and harvested at the time points indicated. Proportion of mitotic cells was determined by phospho-Histone H3 (S10) staining (mean ± SEM; n = 3 biological repeats; Student’s t test (two-sided, equal variance): **p = 0.0023). Data on bar graph underneath represents the 90 min time point (mean ± SEM; n = 3 biological repeats; Student’s t test (two-sided, equal variance): **p = 0.0023). Uncropped blots, raw and processed graphical data provided as a Source Data file.

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