Extended Data Fig. 2: DNA damage in micronuclei in synchronized and unsynchronized cells depends on the catalytic activity of APE1.
From: Breakage of cytoplasmic chromosomes by pathological DNA base excision repair
(a) APE1-AID-SMASh degradation in unsynchronized RPE-1 cells. Control: no drugs; -APE1: 12 h drug treatment prior to western (Dox, IAA and ASV). GAPDH is the loading control. (b) MN γ-H2AX FI with or without APE1 degradation in unsynchronized RPE-1 cells (mean with 95% CI, ****P < 0.0001, Two-tailed Mann-Whitney U test, control: n = 133; -APE: n = 123). (c) Aggregated data from three replicates for MN DNA damage after APE1-AID-SMASh degradation in unsynchronized RPE-1 cells. Drugs to induce APE1-AID-SMASh degradation were added 12 h prior to treatment with MPS1i to generate MN. After 48 h cells were fixed and DNA damage was assessed (mean with SD, ***P = 0.0003, two-tailed unpaired t-test, control: n = 379; -APE1: n = 353). (d) Aggregated data from three replicates for the example in Fig. 1g (mean with SD, NS: P = 0.3606, ***P = 0.0002, Ordinary one-way ANOVA with Dunnett’s multiple comparisons test, control: n = 424; APE1 add-back: n = 392; APE1-CI add-back: n = 400). (e) MN γ-H2AX FI as shown in Fig. 1g but comparing labeling in EdU+ and EdU- cells for the indicated samples (mean with 95% CI, ****P < 0.0001, Ordinary one-way ANOVA with Tukey’s multiple comparisons test. N values from left to right: n = 53, 288, 41, 177, 71 and 280).
