Fig. 2

TTF1 gene knockdown in ND7–23 cells downregulated the expression of Kiss1 and GnRH and decreased TTF1 binding to the Kiss1 promoter. a ND7–23 cells were infected with LV-TTF1-shRNA for 72 h. EGFP (green) was measured with immunofluorescence microscopy. Optical microscopy was used to obtain bright field images. b TTF1, Kiss1 and GnRH mRNA levels were analysed with qRT-PCR after transfection with LV-TTF1-shRNA. Cells transfected with LV-EGFP were used as a negative control. c TTF1 protein concentration was determined by western blotting analysis. d The effect of TTF1 on the activity of the Kiss1 promoter was detected with a Dual-Luciferase Reporter Assay. TTF1-KD, TTF1-knockdown group. TTF1-OE, TTF1-overexpression group. NC, lentivirus negative control group. Scale bars, 200 μm a. The results are shown as the mean ± S.E.M. **P < 0.01