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. 2014 Jun 9;205(5):663–675. doi: 10.1083/jcb.201311050

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© 2014 Broering et al.

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Figure 2. — BRCA1 amplifies ATR and TOPBP1 on unsynapsed sex chromosomes. (A) Immunostaining of meiotic chromosome spreads of adult meiosis using anti-SYCP3, -γH2AX, and -ATR antibodies. (B and C) Percentage of pachytene spermatocytes exhibiting amplified ATR accumulation in first wave meiosis (B) and adult meiosis (C). (D) Immunostaining of meiotic chromosome spreads using anti-SYCP3 and -TOPBP1 antibodies. (E and F) Percentage of pachytene spermatocytes exhibiting amplified TOPBP1 accumulation in first wave meiosis (E) and adult meiosis (F). (G) Immunoprecipitation using the anti–phospho-ATM antibody, followed by detection of coimmunoprecipitation using ATM, ATR, and TOPBP1 antibodies. 10-fold more material was used in the immunoprecipitation than is present in the input. The migration of the nearest protein molecular mass size markers are shown. Images in the same row are from different parts of the same gel. Arrows indicate sex chromosomes. Arrowheads indicate ectopic foci on autosomes. P-values were derived from an unpaired t test. **, P < 0.001. Bars, 10 µm.