Fig. 3

Igf2bp3 is a target for miR-6240. A Potential miRNAs targeting Igf2bp3. B, C RT-qPCR showing the expression of candidate miRNAs in C2C12 (B) and mouse TA muscle (C). D RT-qPCR showing the expression pattern of miR-6240 in different mouse tissues. E Diagram showing the WT/MT reporter constructs that are utilized to pinpoint the Igf2bp3 3' UTR’s miR-6240 target areas. F The relative ratios of firefly luciferase (FL) to renilla luciferase (RL) were ascertained 24 h following the transfection of the plasmids into C2C12 myoblasts. G The plasmids shown were co-transfected with miR-6240 mimics into HEK-293T cells; they were cultured for 24 h, whereupon the relative FL/RL ratios were calculated. H The plasmids shown were co-transfected with either control miRNA or miR-6240 mimics into C2C12 myoblasts; they were cultured for 24 h, and then the relative FL/RL ratios were determined. I, J The expression of both Igf2bp3 mRNA (I) and protein (J) were downregulated by miR-6240 mimics. K, L The RIP experiment's enrichment of miR-6240 (K) and Igf2bp3 (L). GAPDH or U6 was utilized as an internal control. N = 3 in each group. Data are represented as mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ns, not significant (Student’s t-test)