Fig. 4.
Levels of protein translation and MPF activity affect the process of meiotic resumption in Dcaf13-deleted oocytes. a HPG fluorescent staining results showing protein synthesis activity in oocytes at the indicated stages. Oocytes were isolated from 4-week-old Dcaf13fl/fl and Dcaf13fl/fl;Gdf9-Cre mice, and incubated in M2 medium containing 50 μM HPG for 1 h prior to staining. Total numbers of oocytes used (n) in each stage are indicated. Scale bars, 50 μm. b Quantification of HPG signal intensities in a. **P < 0.01, ***P < 0.001, by two-tailed Student’s t tests. Error bars, SEM. c Western blot results showing levels of CDC25B and cyclin B1 in oocytes before and after GVBD. Samples of 100 oocytes were collected before and after culture for 4 h. DDB1 was blotted as a loading control. d, e GVBD rates of cultured Dcaf13fl/fl and Dcaf13fl/fl;Gdf9-Cre oocytes that were microinjected with mRNA encoding cyclin B1 and CDC25B or Flag-vector mRNA (as a control) at the GV stage. The microinjected oocytes were incubated in M16 medium containing milrinone for 10 h and then further cultured in milrinone-free medium for 6 h. **P < 0.01 by two-tailed Student’s t tests. ns non-significant. Error bars, SEM