Fig. 2

EDC4 is required for genome maintenance. a Immunoblots showing the inhibition efficiency in HeLa cells of FANCD2 and EDC4 by three different siRNA sequences used. Two replicas are shown for inhibition. b Inhibition of EDC4 reduces the survival of HeLa cells exposed to MMC. Data shown represent results from five independent experiments. Error bars indicate mean ± s.d. Means were statistically compared using the two-tailed Student’s t test. Statistical analysis comparing the means of the Mock-treated vs. siRNA-treated samples were performed with the following p values: siFANCD2 (10 nM, p = 0.0376; 25 nM, p = 0.0114), siEDC4-1 (10 nM, p = 0.0747; 25 nM, p = 0.0018), siEDC4-2 (10 nM, p = 0.0227; 25 nM, p = 0.0199), and siEDC4-3 (10 nM, p = 0.2004; 25 nM, p = 0.0028). c EDC4 depletion causes chromosome fragility after DEB exposure in primary fibroblasts. Data shown represent results from the analysis of 40 metaphases (two experiments of 20 metaphases each one). Error bars indicate mean ± s.e.m. The statistical test performed in each inhibition to compare breaks per cell (0 vs. 0.1 µg/mL DEB) was a Mann–Whitney test. d CRISPR/Cas9-generated EDC4−/− HEK293T cell line shows a decrease in survival after DEB treatment that is reversed when the WT EDC4 is expressed. Data shown represent results from four combined independent experiments. Error bars indicate mean ± s.d. Means were statistically compared using the two-tailed Student’s t test. Statistical analysis comparing the means of the WT cell line with the samples were performed with the following p values: FANCQ−/− (p = 0.001; p = 0.001), EDC4−/− (p = 0.000; p = 0.000), and EDC4 corrected (p = 0.613; p = 0.173)