Figure 3.

Regulation of PER2 in decidualizing HESCs. A) Primary HESC cultures were treated with 8-br-cAMP, MPA, or a combination for 48 hours and PER2 transcript levels measured. The data show relative change (mean ± sem) in mRNA levels compared to vehicle-treated undifferentiated cultures established from 3 different biopsies. B) Primary cultures remained undifferentiated or were decidualized for 48 hours prior to treatment with 5 μg/ml actinomycin D. RNA was extracted at the indicated time points and subjected to qRT-PCR analysis. C) Binding of CLOCK to E2, a noncanonical E-box enhancer in the PER2 promoter, was assessed by ChIP in 3 independent primary cultures, either undifferentiated (0 hour) or decidualized for the indicated time points. The data show relative enrichment compared to input. D) In parallel, CLOCK binding to a regulatory E-box element (E5) in the PER1 promoter was determined. The data show the mean ± sem. **P < 0.01; ***P < 0.001.