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. 2020 Jul 1;219(8):e201907016. doi: 10.1083/jcb.201907016

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© 2020 Sofroni et al.

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Figure 8. — Characterization of CYCB3;1 in meiosis. (A) Confocal laser scanning micrographs showing the localization of a functional reporter for CYCB3;1 (CYCB3;1:GFP in green) throughout meiosis. Scale bar, 10 µm. (B) Confocal micrographs of TagRFP:TUA5 (magenta) and CYCB3;1:GFP (green) at late prophase and metaphase I and II (from Video 7). CYCB3;1 colocalizes with the first but not the second spindle. Scale bar, 20 µm. (C) CYCB3;1 forms a complex with CDKA;1. Pull-down assay using Strep-CDKA;1 in the presence or absence of HisMBP-CYCB3;1. The input and pull-down fractions were detected by immunoblotting with anti-Strep (upper panel) and anti-MBD (lower panel) antibodies. (D) Chromosome spread analysis of cycb3;1/− versus VF cdka;1/− cycb3;1/− during metaphase I, anaphase I, telophase I, interkinesis, metaphase II, and telophase II. Red arrows mark lagging chromosomes at anaphase I and telophase I. Scale bar, 10 µm. (E and F) Confocal laser scanning micrographs of TagRFP:TUA5 during metaphase I in cycb3;1/− (E) and VF cdka;1/− cycb3;1/− (F). Microtubule arrays are altered in VF cdka;1/− cycb3;1/− as represented by irregular spindles at metaphase I. Scale bar, 10 µm.