Figure 3.

Bcl2l10 regulates the expression of Tpx2, Aurka, and phosphorylated Aurka. (A) Tpx2 RNAi treatment resulted in specific suppression of Tpx2 mRNA expression. In addition, Aurka mRNA was decreased by Tpx2 RNAi. Oocyte-specific H1foo was used as an internal control. (B) Experimental design for Bcl2l10 RNAi experiment. After oocytes were microinjected with Bcl2l10 dsRNA, intact GV oocytes were incubated for 8 h in IBMX-supplemented M16 medium for Bcl2l10 degradation followed by further culture for 16 h in plain M16 medium for in vitro oocyte maturation. Because Bcl2l10 RNAi oocytes were arrested at the MI stage, oocytes of each group were collected at the indicated time points for MI and used for Western blot analysis. (C, D) Tpx2 and Aurka protein expression decreased, whereas T288-Aurka increased after Bcl2l10 RNAi treatment. The lysate of 200 oocytes was loaded in each lane. α-Tubulin was used as a loading control. The experiment was performed 3 times, and the data are presented as the mean ± SEM. Asterisks indicate statistical significance at p < 0.05. Control: non-injected MI oocytes; GFP RNAi: normal MI oocytes after GFP RNAi was used as non-targeting control for RNAi experiments; Bcl2l10 RNAi: MI-arrested oocytes after Bcl2l10 RNAi treatment.