Figure 3
From: 14-3-3 epsilon prevents G2/M transition of fertilized mouse eggs by binding with CDC25B
Overexpression of 14-3-3ε and CDC25B mutants induces mitotic resumption. A: Western blot analysis of CDC25B and 14-3-3ε expression at 7 h after various 14-3-3ε mRNA and Cdc25b mRNA microinjection. Eggs were either not microinjected or microinjected with TE buffer as negative control groups and eggs injected with Cdc25b-S321A mRNA or Cdc25b-WT mRNA solely as the positive controls. Western blotting was performed using CDC25B, 14-3-3ε, phospho-WEE1B-Ser15 and β-actin antibodies in different microinjected groups. 200 eggs are loaded onto each lane. B: The cleavage rate in cultured mouse embryos 12 h after various Cdc25b mRNA and 14-3-3ε mRNA injection in the presence of 2 mmol/l dbcAMP. A x2 test was used to evaluate the differences between multiple experimental groups. Bars represent means ± S.D of three independent experiments and 300 eggs are calculated for each group. C, D: MPF activity was detected at indicated time points in various mRNAs injection and control groups by scintillation counting (C) and autoradiography (D). One-way analysis of variance followed by a Least Significant Difference (LSD) test was used to evaluate the differences in the MPF activity assay. Bars represent means ± S.D of three independent experiments. E: Western blot analysis of phosphorylation status of CDC2-Tyr15. 200 eggs are loaded onto each lane.