FIG. 6.

The specific MMP2/9 V inhibitor prevents steroid production in primary mouse granulosa cells. Primary mouse granulosa cells were treated with LH or LH plus MMP2/9V inhibitor (20 μM). Progesterone levels (A) in the medium were measured by RIA and amphiregulin levels in concentrated medium were measured by ELISA (B). The mRNA levels of amphiregulin or epiregulin were measured by real-time PCR (C). Data are represented as fold induction over MOCK or inhibitor alone (MMP2/9V). Fluorogenic peptide substrate III was used to determine the effect of the MMP2/9V inhibitor on ADAM17 activity in granulosa cell lysates (D) and fluorogenic peptide substrate IX was used to determine the effect of the same inhibitor on MMP activity using purified MMP2 and MMP9 (E). Data are represented as fold induction over untreated control (CON) (D) or fold induction over MMP2 and MMP9 (E). All experiments were performed at least twice with identical results. Each bar represents the average ± SEM. *P < 0.05 relative to MOCK or MMP2/9V alone; **P < 0.05 relative to LH using Student two-tailed t-test.