Fig. 2

CIT-K knockdown reduces BRCA1 levels independently from proliferative state.

a, g Schematic representation of the experiments performed. Quantification of the percentage of EdU positive ONS-76 cells 24 h (b) or 48 h (h) after transfection of the indicated siRNAs. Representative images of ONS-76 cells analyzed 24 h (c) or 48 h (i) after siRNAs transfection and treatment with 10 μM EdU for 30’, to label cells in S-phase. Cells were immunostained for BRCA1 and nuclei were counterstained with DAPI. Scale bars: 10 μm. d, j Quantification of BRCA1 foci per nucleus, in samples treated as in (c, i), respectively. e, k Quantification of BRCA1 foci per EdU-positive nucleus, in samples treated as in (c, i), respectively. f, l Quantification of BRCA1 foci per EdU-negative nucleus, in samples treated as in (c, i), respectively. All immunofluorescence quantifications were based on at least four independent biological replicates; >300 cells were analyzed per condition in each replicate. Error bars, SEM. *P < 0.05, ****P < 0.0001; unpaired two-tailed Student’s t-test for EdU positive cells; Mann– Whitney U test for foci.