Fig. 4

KLF4 is the miR-206 target gene, functioning as an activator of adipocyte differentiation. a Overlap of three miRNA target bioinformatic prediction algorithms. b The potential binding site of miR-206 in the KLF4 mRNA 3′-UTR. c The potential binding site (red) of miR-206 in the KLF4 mRNA 3′-UTR is highly conserved among vertebrates. d Compare the fold change of KLF4 in MYOD1^OE, MYOD1^NC, and MYOD1^KO cells at day 5 compared to day 0. e, f Dual-luciferase reporter assay indicated that miR-206 could bind to the predicted binding site of the KLF4 mRNA 3′-UTR. g, h miR-206 over-expression inhibited KLF4 mRNA and protein expression in ICPs. i, j miR-206 inhibition promoted KLF4 mRNA and protein expression in ICPs. k, l KLF4 mRNA and protein levels at different time points during ICPs differentiation. m Over-expression of KLF4 could counteract the inhibition effect of miR-206 over-expression on adipocyte differentiation. n mRNA levels of adipocyte marker genes were analyzed with qPCR. Data are shown as mean ± SD of three biological replicates. The t-test was used to analyze the statistical differences between groups. *, P < 0.05