Effects of HIF-1α decoy ODNs transfection on ET-2 mRNA, HIF-1α expression and HIF-1 binding activity in response to hCG in granulosa cells. Animals were sacrificed after 48 h PMSG-treatment for collecting ovaries and granulosa cells were isolated for in vitro culture experiments with hCG and dsODN. (A) Representative ECL gel documents of Western blot analyses depicting the protein level of HIF-1α. (B) Summarized intensities of HIF-1α blot normalized to control. (C) The relative mRNA levels of ET-2 by real-time RT-PCR analysis. (D) HIF-1 binding assay in the nuclear extracts from different granulosa cell treatment groups. Each value represents the mean ± SE. One-way analysis of variance (ANOVA) was used to analyze the data. The asterisk denotes significant values (P < 0.05) by Tukey's multiple-range test. n = 6 batches of cells.