Figure 3
Binding activities of C/EBPβ and HIF1α to the Vegf promoter region in rat GCs undergoing luteinization. (A) Location of the binding sites for transcription factors. Primers for ChIP assay were designed to surround the C/EBPβ binding site (−1148 bp to −1024 bp) and the HIF1α binding sites (−976 bp to −857 bp; region-1, −724 bp to −645 bp; region-2, −470 bp to −369 bp; region-3). Binding activities of C/EBPβ (B) and HIF1α (C) before (0 h) and 12 h after hCG injection were analyzed by ChIP assay. GCs from three rats were pooled at each time point to use for ChIP assay. ChIP assay was performed with antibodies to C/EBPβ, HIF1α and control IgG. The relative binding activities of C/EBPβ and HIF1α were assessed by qPCR. The ratio of IP DNA to INPUT DNA sample was calculated. Data were shown as a ratio of those of 0 h. Each value represents the mean ± SEM of 3 independent experiments. a, P < 0.05 vs. 0 h. PCR products from IP DNA and INPUT DNA were also electrophoresed. The images of representative gels are shown.
