Fig. 2: dTAG-13-mediated reduction of endogenous REC8 in REC8−FKBP12F36V−mClover3 oocytes rapidly recapitulates premature sister chromatid separation events associated with female reproductive aging.
From: A versatile cohesion manipulation system probes female reproductive age-related egg aneuploidy
a, Endogenous REC8 (labeled with GFP antibody) and metaphase I chromosomes (Hoechst) in DMSO-treated or dTAG-13-treated oocytes isolated from REC8−FKBP12F36V−mClover3 mice. Single confocal sections spaced 1 µm apart are shown. b, Quantification of REC8 (labeled with GFP antibody) fluorescence intensity in DMSO-treated or dTAG-13-treated metaphase I oocytes isolated from REC8−FKBP12F36V−mClover3 mice. c, Images from representative time-lapse movies of sister chromatids (H2B−mScarlet) in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12F36V−mClover3 oocytes. t = 0 minutes denotes the start of live imaging experiment. d, Quantification of the number of single chromatids per egg at 90 minutes of live imaging in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12F36V−mClover3 oocytes. e, Quantification of the proportion of cells containing single chromatids in DMSO-treated or dTAG-13-treated eggs matured in vitro from REC8−FKBP12F36V−mClover3 oocytes. f, Measurements of chromatid scattering volumes from time-lapse movies of DMSO-treated or dTAG-13-treated metaphase II-arrested eggs matured in vitro from REC8−FKBP12F36V−mClover3 oocytes. Lines represent mean values, and error bars indicate s.e.m. Data are from three independent experiments. Statistical significance was evaluated using Student’s t-test (b,d,e) or two-way ANOVA (f). The number of analyzed oocytes is indicated in brackets and italics.
