(A) Expression analysis of DLX3 and miRNAs in sheep skin and SFFs by real-time RT-PCR. The data were normalized against GAPDH (gene expression, left panel) and 5S ribosomal RNA (miRNA expression, middle and right panels), respectively. All PCRs were performed in triplicate; the data are expressed as the mean ± S.D. of three independent experiments. (B) Effects of miR-31 on DLX3 3'UTR luciferase reporter activity in SFFs and HaCaT cells, respectively. The graph is a mean of the three haplotypes (CCGI, TTAD or TCAD). About >30% suppression in the reporter activity was observed, compared to the control (cells cotransfected with psiCHECK2 vector and miR-31 mimic). (C, D) Effects of miR-188 mimic on DLX3 3'UTR reporter activity of psiCHECK2-TCAD and psiCHECK2-TCAI reporters in SFFs and HaCaT cells. MiR-188 mimc reduced the reporter activity of psiCHECK2-TCAD reporter by over 50%, but did not alter the reporter activity of psiCHECK2-TCAI reporter. NC: miRNA mimics negative control. EV: psiCHECK2 empty vector. (E, F) Effects of miR-3957-5p mimic on the reporter activity of psiCHECK2-TCGI and psiCHECK2-TCAI reporters in SFFs and HaCaT cells. MiR-3957-5p mimic caused no significant changes in the reporter activity of both psiCHECK2-TCGI and psiCHECK2-TCAI reporters. *p < 0.05; **p < 0.01.