Fig. 5

CCNB3 mutations affect the activity of CDK1 and APC/C. (A, D) A diagram depicting flow of mouse GV injection. GV oocytes were injected with WT or mutant CCNB3 cRNA into cytoplasm and subsequently cultured for 3 or 10 h, following collecting the oocytes. (B-C) Western blot analysis of oocytes injected with cRNA and cultured for 3 h in order to assay the CDK1 activity by testing the phosphorylation of Lamin A/C and PP1-α. (E, H) Western blot analysis of oocytes injected with cRNA and cultured for 10 h in order to assay the CDK1 activity by testing the phosphorylation of Lamin A/C and PP1-α. (F, G) Western blot analysis of oocytes injected with cRNA and cultured for 10 h in order to trace protein level changes of the APC/C substrates, securin and Ccnb1 protein, and the level of its activators, CDH1 and CDC20. (I) Relative CDH1, CDC20, CCNB1 and securin level after 10 h culture. Data are shown as the means ± SEM. For each kind of variants,n = 3 biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001, **** p < 0.0001