RNF2 aggregation correlates with loss of 53BP1 response foci and depletion of KDM2A/B rescues 53BP1 response foci formation at sites of damage. (A) Representative immunofluorescence images of 53BP1 signal (green) and RNF2 signal (magenta) in −dox, +dox, +eto, or +dox +eto iDUX4 cells (scale bar = 20 μm). White arrows indicate nuclei that contain RNF2 aggregates but no 53BP1 signal/recruitment to sites of damage. Asterisks indicate nuclei with pan-nuclear RNF2 signal and 53BP1 foci. Images are representative of three independent experiments conducted on separate days. (B) Fraction nuclei with RNF2 signal type and 53BP1 recruitment to sites of damage. RNF2 pan-nuclear signal (pan) or RNF2 aggregates (foci) in +dox only or +dox +eto iDUX4 cells. Nuclei are indicated for each independent experiment and N ≥ 100 nuclei per condition. (C) Expanded data from Fig. 7 D. Fraction total nuclei with either γH2A.X signal only or both γH2A.X and 53BP1 signal in +dox cells with (siKDM2A/B) or without (siCTRL) KDM2A/B depletion in nuclei that contained no HSATII RNA foci (HSATII−) or contained HSATII RNA foci (HSATII+). N ≥ 100 nuclei between all fields; dots indicate the average between fields. (B) Data represent means ± SD. Statistical differences between groups were analyzed with one-way ANOVA Tukey’s multiple comparison test between each group and a control.
Figure S4.

RNF2 aggregation correlates with loss of 53BP1 response foci and depletion of KDM2A/B rescues 53BP1 response foci formation at sites of damage. (A) Representative immunofluorescence images of 53BP1 signal (green) and RNF2 signal (magenta) in −dox, +dox, +eto, or +dox +eto iDUX4 cells (scale bar = 20 μm). White arrows indicate nuclei that contain RNF2 aggregates but no 53BP1 signal/recruitment to sites of damage. Asterisks indicate nuclei with pan-nuclear RNF2 signal and 53BP1 foci. Images are representative of three independent experiments conducted on separate days. (B) Fraction nuclei with RNF2 signal type and 53BP1 recruitment to sites of damage. RNF2 pan-nuclear signal (pan) or RNF2 aggregates (foci) in +dox only or +dox +eto iDUX4 cells. Nuclei are indicated for each independent experiment and N ≥ 100 nuclei per condition. (C) Expanded data from Fig. 7 D. Fraction total nuclei with either γH2A.X signal only or both γH2A.X and 53BP1 signal in +dox cells with (siKDM2A/B) or without (siCTRL) KDM2A/B depletion in nuclei that contained no HSATII RNA foci (HSATII−) or contained HSATII RNA foci (HSATII+). N ≥ 100 nuclei between all fields; dots indicate the average between fields. (B) Data represent means ± SD. Statistical differences between groups were analyzed with one-way ANOVA Tukey’s multiple comparison test between each group and a control.

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