Fig. 4

miR-339 activates CYP19A1 transcription in sow GCs by acting as an endogenous saRNA. a miR-339 does not influence the activity of CYP19A1 promoter with the miR-339 MRE was mutated. A reporter vector of the mutant CYP19A1 promoter containing the MRE motif of miR-339 was generated, and co-treated with miR-339 mimics, and luciferase activity was measured. n = 3. b miR-339 activates CYP19A1 transcription in GCs. qPCR was performed to detect the CYP19A1 mRNA levels in GCs after treated with miR-339 mimics for 24 h, and 48 h. GAPDH was used as an internal reference. n = 3. c-d miR-339 increases CYP19A1 protein levels in GCs. Western blotting was performed to detect the CYP19A1 protein levels in GCs after treated with miR-339 mimics for 24 h (c), and 48 h (d). GAPDH was used as an internal reference. n = 3. e–g ChIP assays. Specific primers for amplifying CYP19A1 promoter containing miR-339 MRE motif are designed (e) to amplify DNA products enriched by specific antibodies against AGO2 (f), and H3K4me and H3K9ac (g) in GCs. F, forward primer. R, reverse primer. Values are means ± SEM. *, P < 0.05; ns, no significant