Figure EV5. PIDD1 localization to DAs is required for PIDDosome activation in response to DNA damage also in RPE1 cells.

1. Dot plot showing the average pixel intensities of PIDD1 at individual parent centrioles in A549 cells of the indicated genotypes and treatments (CPT = camptothecin, ZM = ZM447439). Mean values (red lines) ± s.e.m. are reported. N > 50 centrosomes were assessed for each condition in as many individual cells; a.u. = arbitrary units. Kruskal–Wallis test (n.s. = non‐significant).
2. RPE1 cells of the indicated genotypes were treated either with CPT or ZM or DMSO for 24 h. Samples were subjected to immunoblotting; n = 3 independent experiments.
3. RPE1 cells treated as in (B) were subjected to fluorescence microscopy and centrosome abundance was assessed by counting the number of γ‐tubulin‐positive centrioles per cell across different genotypes and treatments. Mean values ± s.e.m. are reported. N = 3, ≥ 50 cells from each independent experiment. ANOVA test (n.s. = non‐significant).
4. Dot plot showing the average pixel intensities of PIDD1 at individual parent centrioles in RPE1 of the indicated genotypes and treatments. Mean values (red lines) ± s.e.m. are reported. N > 50 centrosomes were assessed for each condition in as many individual cells; a.u. = arbitrary units. Kruskal–Wallis test (n.s. = non‐significant).
5. RPE1 cells of the indicated genotypes were either left untreated or treated with Nutlin‐3a (3.3 µM or 10 µM). Cells were subjected to immunoblotting; n = 3 independent experiments.
6. A549 cells were either treated with 10 µM Nutlin‐3a for 24 h or left untreated. Cells were subjected to fluorescence microscopy and centrosome abundance was assessed by counting the number of γ‐tubulin‐positive centrioles per cell across different genotypes and treatments. Mean values ± s.e.m. are reported. N = 3, ≥ 50 cells from each independent experiment. ANOVA test (n.s. = non‐significant).

Source data are available online for this figure.