Fig. 2

CCDC6 depletion affects γH2AX foci formation and impairs Homologous Recombination repair. A Immunofluorescence images showing, compared to untreated control cells (a,f,k), the impact of CCDC6 silencing, followed by exposure to PARPi [1 μM] (b,c,g,h,l,m) or PARGi [1 μM] (d,e,i,j,n,o) for 48 h, on γH2AX nuclear foci formation in Kuramochi, OVCAR3 and OV-90 cells. Scale bar 50 μm. B Graphs represent the percentage of cells with more than 15 foci. Error bars indicate the standard error mean derived from three independent experiments. Statistical significance was verified by 2-tailed Student's t-test (* p < 0.05; ** p < 0.01 and *** p < 0.001). C Kuramochi, OVCAR 3 and OV-90 cells, after pre-treatment either with vehicle or P5091 [2.5 μM] for 4 h, were transfected with DR-GFP alone, as control, with HA-ISceI (ISceI) and with both HA-ISceI (ISceI) and Myc CCDC6 plasmids. The percentages of GFP positive cells, compared to controls, were plotted as histograms, representative of the mean of three independent experiments. Error bars indicate the measurement of the standard error mean. Statistical significance was verified by 2-tailed Student's t-test (* p < 0.05; ** p < 0.01 and *** p < 0.001). D, E, F Myc CCDC6 and HA-ISceI proteins expression were assessed by anti-Myc and anti-HA antibodies at Western Blot. Anti-Tubulin immunoblots are shown as loading control