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. 2021 Sep 24;9:666303. doi: 10.3389/fcell.2021.666303

FIGURE 5.

FIGURE 5

Regulation of NR5A2 activity by LGR4 mediated canonical WNT/CTNNB1 signaling pathway. (A) The mRNA levels of Cyp11a1 and Hsd3b1 in the WT and LGR4 mutant oviducts at the estrus stage. (B) Diagram of conserved NR5A1/2 binding sites in the Cyp11a1 and Hsd3b1 promoters predicted in silico. (C) IHC assays to detect the NR5A2 protein location in the WT or Lgr4-deficient oviducts. (D) IHC assays to detect the CTNNB1 protein level in the WT or Lgr4-deficient oviducts. (E) The mRNA levels of WNT target genes in WT and LGR4 mutant oviducts at estrus stage. (F) The chromatin of primary oviductal epithelial cells from WT or Lgr4 mutant mice was immunoprecipitated with a NR5A2 antibody. Immunoprecipitated DNA was amplified with the sets of specific primers for the indicated NR5A2 binding sites in Cyp11a1 and Hsd3b1 promoters (upper panel). The relative fold change of Cyp11a1 and Hsd3b1 genes of the primary oviductal epithelial cells after the treatment of WNT4/RSPO2 or CT99201 (lower panel) are presented. ∗p ≤ 0.05; ∗∗p ≤ 0.01; ∗∗∗p ≤ 0.001. Scale bar, 50 μm in (C,D). IHC, immunohistochemistry.