Fig. 5

Functional analyses of the NR5A1 mutant. A NR5A1 mRNA levels in 293 T cells according to qRT-PCR analysis: non-transfected (Control) and transfected with an empty vector (Vector), Myc-tagged WT (WT), or c.64G > T (p.G22C) NR5A1-Mut (Mut) vectors. B NR5A1 expression in 293 T cells according to western blot analysis (same grouping as A, the raw figures in Supplementary Fig. 3A&B.). C Nuclear localization of the NR5A1 mutant according to immunocytochemical analysis (same grouping as A). Scale bar, 50 μm. D EMSAs results showing altered DNA binding by the NR5A1 mutant. Nuclear extracts were prepared from four groups of cells (same grouping as A, the raw figures in Supplementary Fig. 3C). E Transcriptional activity of the NR5A1 mutant. Dual-luciferase activity detected in cells co-transfected with the WT plasmid and AMH reporter or the NR5A1-Mut plasmid and AMH reporter. The internal fluorescence reference was pRL-TK Renilla luciferase; t-test was applied for.^*p < 0.05