Fig. 4

miR-338-3p overexpression suppressed the E₂-production capacity of primary human granulosa cells. (A) The GCs were inoculated in 6-well plates and expanded by routine culture for 24 h. Next, the original culture medium was replaced with cell-starvation medium, followed by 24 h of further culture. The cells were then transfected with miR-338-3p mimics/inhibitor to manipulate the expression level of miR-338-3p and cultured for 48 h, after which the culture supernatant was collected to determine the E₂ levels by performing an electrochemiluminescence immunoassay. (B) Primary human GCs were transfected with miR-338-3p mimics/inhibitor to manipulate miR-338-3p expression, the cells were cultured for 48 h, and western blotting analysis was performed to determine the expression level of aromatase. (C) Image J software was used to perform grayscale analysis of the aromatase protein bands. The bar chart displays the ratio of the grayscale value of the aromatase protein to that of the GAPDH protein. *P < 0.05, **P < 0.01, ***P < 0.01