Fig. 4: DNA damage-induced PIDD1 phosphorylation triggers SUMOylation. | Nature Communications

Fig. 4: DNA damage-induced PIDD1 phosphorylation triggers SUMOylation.

From: Stepwise phosphorylation and SUMOylation of PIDD1 drive PIDDosome assembly in response to DNA repair failure

Fig. 4

a, b PIDD1—/— HeLa cells transfected with indicated FLAG-PIDD1 variants, treated with MMC and Go6976 with or without SUMO E2 ligase inhibitor 2-D08 were harvested at 24 h and analyzed by western blot. c Pidd1—/— MEF transfected with indicated FLAG-PIDD1 variants and treated with or without MMC and Go6976 were harvested at 24 h, immunoprecipitated with anti-FLAG antibody and analyzed by western blot. d PIDD1—/— C2.Pro-BiFC HeLa cells were transfected with indicated FLAG-PIDD1 variants, treated with or without MMC and Go6976 (5 μM each) and imaged at 24 h. At least 40 cells per sample were scored in each of n = 3 independent experiments. Representative images were quantified (right), with data expressed as means +/− SD. **p < 0.005; ***p < 0.001; ns, non-significant, two-tailed Student’s t-test. e, f PIDD1—/— HeLa cells transfected with indicated FLAG-PIDD1 variants and treated with MMC and Go6976 (e) with or without ATR inhibitor BAY-1895344 (BAY, 0.75 mM) (f) were harvested at 24 h, immunoprecipitated with anti-PIDD1pT788 antibody and analyzed by western blot. Cells in (a–c, e, f ) were TdR-synchronized as described in Supplementary Tables 1B and 1C. Unless otherwise indicated, drugs were given at 1 μM. Source data are provided as a Source data file.

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