Fig. 3.

Bime2 localization. a HA-tagged Bime2 (red) localizes to the chromatin of meiotic nuclei from the start of nuclear elongation to the end of prophase. It is undetectable in metaphase I. (Numbers refer to the stages in Fig. 1a.) b Similar to Dmc1, Bime2 localization is Spo11 dependent. c Bime2 foci are not formed in Dmc1-depleted cells. Preparations were co-stainined for Dmc1—the absence of Dmc1 in meiotic germline nuclei confirms the efficiency of RNAi-mediated Dmc1 knockdown. Foci seen in somatic nuclei represent antibody cross-reactivity with Rad51. For Dmc1 localization in wild-type cells, see Fig. 2e. d Co-staining of Bime2-HA and Dmc1. Although both proteins form foci on the chromatin of meiotic prophase nuclei, they do not colocalize. Chromatin was stained with DAPI (blue). The bloated appearance of nuclei is caused by detergent treatment to remove non-chromatin-bound proteins. Scale bar: 10 μm