Figure 6

TBP2 degradation takes place after the establishment of transcriptional shut down. (A) A scheme of the experiment shown in (B) is presented. Oocyte nuclei were injected with 0.6 ng of hsp70 and 0.3 ng of Cyr61 promoter constructs. After a delay of 12 hr oocytes were divided into two groups and one group was treated with progesterone (final concentration 2 μg/ml). Oocytes were synchronized at germinal vesicle breakdown (GVBD). Some were processed immediately for chromatin immunoprecipitation (ChIP) and immunoblotting along with control oocytes (not treated with progesterone) whereas others were analyzed 5 hr after GVBD. (B) Recruitment of pol-II (upper graph) and TBP2 (lower graph) to hsp70 and Cyr61 promoters in control oocytes versus oocytes at GVBD and 5 hr after GVBD was assayed by ChIP. Below the graphs expression of TBP2 has been shown corresponding to each sample. TFIIF served as a loading control.