Figure 7.
Suppression of DUOX2 reduces GnRH-induced Fshb mRNA. LβT2 cells were transduced with lentiviral shRNA targeting mouse Duox2 for 2 days. Transduced cells were starved for 16 hours and then stimulated with 10 nM GnRH for 5 hours. A, DUOX2 protein was measured by Western blotting. Both unstimulated 0-hour and 5-hour GnRH-stimulated DUOX2 levels were significantly reduced in comparison to those of control vector-transduced cells. B, Fshb and Lhb mRNA was analyzed by quantitative real-time PCR. Gapdh mRNA was used as an internal control for normalization across samples. Relative transcript levels were determined using the ΔΔCT method. Fshb mRNA was not significantly induced by GnRH in shDUOX2-transduced cells, but Lhb mRNA was unaffected.