Figure 7 |. Loss of centromere identity in the absence of RAD51.

(A) Western blot confirming the depletion of RAD51 in asynchronous HCT116 cells, 72 hours after siRNA transfection. For all Western blots, Lamin-A levels were used as a loading control. (B) Quantification of CENP-A and CENP-B levels in asynchronous HCT116 cells, 96 hours after siRNA-mediated depletion of RAD51. CENP-B foci were used to define centromere loci, and quantification was performed as in Figure S5. (C) Western blot confirming the depletion of RAD51 in asynchronous hTERT-RPE1 cells, 96 hours after siRNA transfection. Dashed line indicates where the blot was cropped. (D) As in (B) but for asynchronous hTERT-RPE1 cells. (E) Western blot confirming the depletion of RAD51 in serum-starved hTERT-RPE1 cells. Cells were serum-starved for 24 hours prior to siRNA treatment and harvested 72 hours later. (F) As in (B) but for serum-starved hTERT-RPE1 cells. (G) Representative images of (F). At least 30 cells were imaged per experimental condition, and the medians of each experimental condition were used to perform a two-sided unpaired t-test (*p<0.05, **p<0.01). (H) Model for the role of RAD51-mediated recombination at spontaneous centromere HOR DNA breaks. See also Figure S8.