Fig. 4

FANCD2-dependent ssDNA formation upon cisplatin treatment. a HeLa cells were transfected with siRNA and labeled with IdU prior to cisplatin exposure for 24 h. After fixation, cells were stained for ssDNA, using an anti-IdU antibody, and for pS33 (RPA2 phosphorylated at Ser33). DAPI was employed to visualize the nuclei. b Quantification of nuclear fluorescence representing ssDNA induced by the indicated treatments (N = 368–594 nuclei from 3 experiments). c Quantification of nuclear fluorescence representing RPA2 phosphorylation at Ser33 (N = 440–580 nuclei from 2 to 3 experiments). In panels b and c, horizontal lines represent median values, boxes 25-75th percentiles and whiskers 10-90th percentiles. *P < 0.05, **P < 0.01, ***P < 0.001 (1-way ANOVA according to Kruskal-Wallis). Scale bar: 10 μm