Fig. 6

Leptin is targeted by miR-874-3p. a Venn diagram of putative miRNA targeting to leptin, and target prediction software miRDB, miRWalk and TargetScan were used for this study. b Predicted binding sites of miR-874-3p within the 3’UTR of leptin mRNA. c and d TW06 and TW02 cells were transfected with miR-874-3p mimics or miR-874-3p inhibitor for 48 h. The leptin mRNA expression levels were examined by QPCR. e Luciferase reporter assays were performed to determine the effect of miR-874-3p on the activity of leptin 3’UTR. f Western blot analysis of leptin and leptin downstream molecules in NPC cells transfected with miR-874-3p mimic, or miR-874-3p inhibitor, and corresponding negative control (NC). g The RIP assay was performed using AGO2 or IgG antibodies in NPC cells and the enrichment of miR-874-3p and leptin were detected by QPCR. h The RIP assays were also performed using AGO2 or IgG to estimate the enrichment of leptin in NPC cells transfected with miR-874-3p mimics and negative control. i The mRNA level of leptin was negatively correlated with miR-874-3p in NPC samples. *P < 0.05, **P < 0.01, ***P < 0.001