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. 2020 Feb 14;9(2):445. doi: 10.3390/cells9020445

Figure 5.

Figure 5

Steroidogenic Factor 1 (SF1) regulates both miR-202-5p and CYP19A1 expression and is associated with estrogen release. (A) Correlation between SF1 and miR-202-5p and CYP19A1. U6 and GAPDH were used as references for miRNA and mRNA expression, respectively. (B) Correlation between E2 in the follicular fluid and the relative mRNA expression of goat GCs. (C,D) Goat GCs were cotransfected with pcDNA3.1-SF1, pcDNA3.1 vector, siRNA-NC and SF1-siRNA, and total RNA was extracted 48 h post-transfection. mRNA levels of miR-202-5p and CYP19A1 were detected by qPCR. (E) SF1 regulates both miR-202 and CYP19A1 promoter activities. HEK293T cells were cotransfected with pcDNA-3.1-SF1 and reporter vectors harboring wildtype or mutant promoters. Renilla and firefly luciferase were assessed and relative luciferase activity was calculated. (F) Chromatin immunoprecipitation (ChIP) assays to detect SF1 binding to pri-miR-202 and CYP19A1 promoters. Experiments were repeated three times. Correlation analysis was performed using Spearman’s rank correlation coefficients. Bars indicate the mean ± SEM of three independent replicates; *p < 0.05, **p < 0.01, ****p < 0.0001.