Fig. 1: MAD2L2 promotes global DNA replication during replicative stress.

a, b Representative images (a) and quantification (b) of survival assays in control (sgCTRL) and MAD2L2-depleted HeLa cells, untreated or treated with 0.5 μΜ MMC or 1 μΜ CIS. Bars represent the mean ± SD. Each dot represents one of three independent experiments. Statistical analysis was performed according to one-way ANOVA with Dunnett’s multiple comparisons test. c, d Assessment of DNA synthesis rates. Schematic with representative images (c) and quantification (d) of EdU incorporation intensity in control (sgCTRL) and MAD2L2-depleted HeLa cells. Cells were incubated with or without 5 μΜ MMC for 24 h, followed by EdU labeling for 2 h before harvesting. Scale bar, 50 μΜ. White bars represent the median of three independent experiments. Statistical analysis was performed according to two-tailed Mann-Whitney test. e Schematic of the SIRF assay, which combines the proximity ligation assay (PLA) with EdU coupled click-iT chemistry to detect the association of target proteins to nascent DNA. Created with BioRender.com. f, g Representative images and quantification of the percentage of cells with MRE11/biotin PLA foci (f) or MAD2L2/biotin PLA foci (g) in HeLa control cells, untreated or treated with 4 mM HU for 2 h. Scale bar, 10 μΜ. Bars represent the mean ± SD. Each dot represents one of three independent experiments. Statistical analysis was performed according to unpaired two-tailed student t-test.