Fig. 8
CFP1 degradation is required for appropriate histone H3 modifications and normal spindle assembly. a Immunofluorescent staining of α-tubulin showing spindle assembly in oocytes microinjected with mRNAs encoding WT or mutated CFP1. Oocyte subcortical actin microfilaments and DNA were labeled by phalloidin (red) and DAPI (blue), respectively. Scale bar, 20 μm. Fully grown GV oocytes from WT mice were used for mRNA microinjection throughout the experiments presented in this figure. b GVBD and PB1 emission rates in oocytes microinjected with mRNAs encoding CFP1 (WT and mutants), as well as the percentages of oocytes containing a normal spindle at MI stage. GV oocytes were microinjected with mRNAs encoding CFP1 (WT and mutants) and were allowed to resume meiotic maturation for 8 h. Error bars, S.E.M. ***P < 0.001 and **P < 0.01 by two-tailed Student’s t tests. n.s., non-significant. c, d Immunofluorescence of CFP1 (c) and H3K4me3 (d) on oocyte chromosome spreads prepared at MI stage. Centromeres and DNA were labeled by CREST immunofluorescence (green) and DAPI staining (blue), respectively. Scale bar, 5 μm. e Quantification of H3K4me3 fluorescent signals in (d). Error bars, S.E.M. ***P < 0.001, **P < 0.01, and *P < 0.05 by two-tailed Student’s t tests. n.s., non-significant. f Chromosome spreads made from MI oocytes overexpressing WT and mutated CFP1. Centromeres, chromosome arms, and DNA were labeled by CREST (green), Topoisomerase II (TOP2, red), and DAPI (blue) staining, respectively. Scale bar, 5 μm
