FIGURE 4.

miR-320 regulates the expression of E2F1/SF-1 and GC function in vivo. A, localization of GFP-labeled miR-320 mimics/mimics NC and miR-320 inhibitors sponge/inhibitors NC lentivirus in the ovaries was examined under a fluorescent microscope with Hoechst staining. Arrows indicate the lentivirus in the GCs of the ovary. B and C, infection efficiency of miR-320 mimics and miR-320 inhibitors sponge virus in GCs (B) and ovaries of mice (C) was determined by real time PCR. Total RNA was extracted from ovaries/GCs and then subjected to analysis to the expression of miR-320, E2f-1, and Sf-1 (C). D, miR-320 regulated the expression of SF-1, E2F1, PCNA, CDK4, CDK2, cyclin D1, caspase-3, and PARP in the ovaries in vivo. Ovaries were infected with lentivirus for 3 days as described above. Expression of SF-1, E2F1, PCNA, CDK4, CDK2, cyclin D1, caspase-3, and PARP in the ovaries was determined by Western blotting. E–G, miR-320 affected steroidogenesis in vivo. Ovaries were infected with lentivirus for 3 days as described above. Sera of the mice were collected for measurement of E₂ (E), progesterone (F), and testosterone (G) levels. B and C and E–G, data represented the mean ± S.E. of three independent experiments performed in triplicate. **, p < 0.01.