Skip to main content
. 2020 Nov 11;11:529757. doi: 10.3389/fgene.2020.529757

FIGURE 3.

FIGURE 3

miR-149-5p suppresses CMTM3 expression and accelerates AR expression by directly targeting the 3′-UTR of CMTM3. (A) The predicted binding site (blue) and mutated site (red) of miR-149-5p in the 3′-UTR of goat CMTM3. (B–D) Dual-luciferase activity assay of the wild-type or mutant 3′-UTR of CMTM3. NC or miR-149-5p mimics were cotransfected with wild-type or mutant CMTM3 3′-UTR luciferase reporters in HEK293T cells (B). Anti-NC or miR-149-5p inhibitors were cotransfected with wild-type or mutant CMTM3 3′-UTR luciferase reporters in HEK293T cells (C). pcDNA3.1(+) or pcDNA3.1(+)-miR-149-5p was cotransfected with wild-type or mutant CMTM3 3′-UTR luciferase reporters in HEK293T cells (D). (E) CMTM3 mRNA expression during hair follicle stem cell proliferation after transfection with NC, miR-149-5p mimics, anti-NC, miR-149-5p inhibitors or pcDNA3.1(+)-miR-149-5p as evidenced by RT-qPCR; total RNA was harvested at 24 h, 48 h, 72 h, and 96 h after transfection. (F) AR mRNA expression during hair follicle stem cell proliferation after transfection with NC, miR-149-5p mimics, anti-NC, miR-149-5p inhibitors or pcDNA3.1(+)-miR-149-5p was evidenced by RT-qPCR; total RNA was harvested at 24 h, 48 h, 72 h, and 96 h after transfection. (G–I) CMTM3 (1:1000 dilution) and AR (1:1000 dilution) protein expression levels were examined after transfection with NC, miR-149-5p mimics (G), anti-NC, miR-149-5p inhibitors (H), and pcDNA3.1(+)-149-5p (I) in GM at 24 h, 48 h, 72 h, and 96 h. The results from each group are shown as the mean ± SEM of three independent replicates. Independent-samples t-tests were used for statistical analysis. Asterisks indicate significant differences. No asterisk means P > 0.05, *P < 0.05, and **P < 0.01.