Figure 5.

miR-24 directly represses Tcf-3 in vitro and in vivo. (a) 3′UTR from mouse and human Tcf-3 indicating the two putative miR-24 binding-sites predicted by the rna22 software. (b) Insertion of mouse Tcf-3 3′UTR decreases luciferase reporter activity. (c) qPCR shows in vitro and in vivo downregulation of Tcf-3 mRNA levels in miR-24-overexpressing systems. (d) Western blot analysis in human primary keratinocytes 48 h post transfection or in P3 transgenic mice showed reduced Tcf-3 protein level. Tubulin was used as loading control. (e) Antago-miR-24 subcutaneous injection in newborn mice significantly reduced endogenous levels of miR-24 in the skin (*P<0.01). (f) Western blot analysis in Antago-24-treated mice showed increased levels of Tcf-3 in mouse skin. Actin was used as loading control