FIG. 9.

Inhibition of protein synthesis at GVBD similarly blocked MPF and Aurora-A reactivation. (A and B) Oocytes were stimulated with progesterone. Individual oocytes were lysed at germinal vesicle, GVBD, or 1 h or 3 h following GVBD. When cycloheximide (CHX, 100 μg/ml) was used, it was added at GVBD. Aliquots of the same extracts were used for MPF assays (A) and immunoblotting with antibodies against Aurora-A (B). Shown is a representative of three independent experiments. (C) Oocytes were either incubated in OR2 (germinal vesicle) or treated with progesterone. Following progesterone treatment, individual oocytes were selected at GVBD and grouped (synchronized within 5 min). Groups of 30 oocytes were either lysed immediately (GVBD, lanes 2 and 4) or further incubated for 3 h in the absence (lane 3) or presence (lane 5) of cycloheximide. Extracts were prepared and subjected to anti-Aurora-A immune kinase assays. Shown is a representative of two independent experiments. MBP, myelin basic protein.