Upregulation of WNT/β-catenin signaling genes in testes of <i>ΔXmiRs</i> mice, and repression of <i>Fzd4</i> by XmiRs.

(A, B) Relative expression of the putative common target genes of miR-871-3p and miR-880-3p (A) and of the downstream genes of WNT/β-catenin signaling (B) in the testes of WT and ΔXmiRs (F2 of the OT84 line) mice at 12 weeks of age was determined by quantitative RT-PCR analysis. The expression in WT testis was set as 1. (C) Potential target sites of miR-871-3p and miR-880-3p in Fzd4-3′-UTR. (D, E) Relative luciferase activity from the reporter vectors with Fzd4-3′-UTR with or without expression vectors for miR-871 (D) or miR-880 (E). Fzd4-3′-UTR ΔmiR-871; Fzd4-3′-UTR with deleted miR-871-3p target site (D). Fzd4-3′-UTR 1–927 or 927–2263 with or without ΔmiR-880; 1–927 or 927–2263 bp fragment of Fzd4-3′-UTR with or without deleted miR-880-3p target sites (E). Luciferase activity was measured 48 h after transfection. Luciferase activity with an empty expression vector was set as 1. Error bars represent standard errors of three biological replicates. *P < 0.05 and **P < 0.01.