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. 2017 Feb 6;114(8):1928–1933. doi: 10.1073/pnas.1615133114

Fig. 3.

Fig. 3.

CENP-A maintains the integrity of the α-satellite repeats. (A) hTERT-RPE1 cells were treated with control siRNA (siCNT) or two different siRNA, #1 and #2 targeting to CENP-A for 72 h. Chromatin-bound proteins were analyzed by Western blotting using anti–CENP-A antibody (Top) and ponceau (Bottom). (B) Examples of aberrant c-CO-FISH patterns in siCENP-A cells using probe set 1. (C) Quantification of aberrant c-CO-FISH. n ≥ 10. Error bars ± SD, **P ≤ 0.005. (D) hTERT-RPE1 lines upon deletion of the CENP-A floxed allele by infection with Adeno-CRE for 6 d. +/+ hTERT-RPE1; +/F one floxed allele; −/F one deleted and one floxed allele. Chromatin-bound proteins were analyzed by Western blotting using anti–CENP-A antibody (Top) and ponceau (Bottom). (E) c-CO-FISH (probe set 2) quantification of centromere instability in CENP-A flox hTERT-RPE1 cell lines as in D; n ≥ 10. Error bars ± SD, ***P ≤ 0.0005. (F) c-CO-FISH of a metaphase spread from cell lines as in D. (Scale bars, 1 μm.) Three centromeres from each figure marked by the colored boxes are zoomed in and shown in the panel Below, as indicated.