Figure 8. The p97–ATX3 complex promotes accurate execution of NHEJ and DSB repair.

1. GFP based reporter assay showing the efficiency of NHEJ repair pathway under indicated siRNA‐depleted conditions in HEK293 cells. Graph represents the quantifications from three independent experiments (*P < 0.05, **P < 0.01, ***P < 0.001; one‐way ANOVA, n = 3, mean + SEM).
2. GFP based reporter assay showing the efficiency of HR repair pathway under indicated siRNA‐depleted conditions in HEK293 cells. Graph represents the quantifications from three independent experiments (^ns P > 0.05, **P < 0.01; one‐way ANOVA, n = 3, mean + SEM).
3. CFA showing the IR sensitivity of 53BP1‐proficient and 53BP1‐deficient MCF7 cells under indicated siRNA‐depleted conditions. Data were collected from two individual experiments with triplicates (^ns P > 0.05; two‐way ANOVA, n = 2, mean ± SEM).
4. CFA showing the IR sensitivity of BRCA2‐proficient and BRCA2‐deficient DLD1 cells under indicated siRNA‐depleted conditions. Data were collected from two individual experiments with triplicates (**P < 0.01, ***P < 0.001; two‐way ANOVA, n = 2, mean ± SEM).
5. Representative IF images showing Rad51 foci in S‐phase (EdU‐positive) U2OS nucleus after 5 h of IR (2 Gy) exposure. Scale bar: 10 μm.
6. Graph represents the quantification of (E) showing average number of Rad51 foci per S‐phase (EdU‐positive) nucleus (^ns P > 0.05, ****P < 0.0001; unpaired t‐test, n = 2, mean + SEM, at least 50 nuclei per condition and experiment).
7. CFA showing the rescue of cell survival in IR‐treated, ATX3‐depleted (siRNA), HeLa cells, after co‐depletion (siRNA) of RNF8 (^ns P > 0.05, *P < 0.05, ***P < 0.001; two‐way ANOVA, n = 3, mean ± SEM).
8. Model of the p97–ATX3 complex in the regulation of RNF8 homeostasis under physiological condition (soluble environment) and in RNF8 chromatin extraction at sites of DSBs to promote NHEJ.

Source data are available online for this figure.