Fig. 7.

NPR3 is a target gene of miR-146a in human adipocytes and murine WAT. a Representative Western blot for NPR3 in SGBS adipocytes overexpressing miR-146a (146a) compared to control cells (Ctrl) with (b) densitometric analysis of 4 independent experiments in relation to β-actin. c Illustration of miR-146a binding sites in the NPR3 3′ UTR predicted by miRWalk 3.0. WT wild type, mut mutated. d Firefly luciferase signal normalized to Renilla luciferase signal of HEK293 cells co-transfected with miR-146a mimic or control (NT) and pmirGLO dual-luciferase plasmid carrying either the two predicted NPR3-miR-146a binding sites (WT), a plasmid with both target sites (site 1 + 2), or a plasmid with either binding site 1 (site 1) or binding site 2 (site 2) mutated. e Npr3 protein expression in gonadal fat pads of miR-146a^−/− (KO) and control (WT) mice after 10 weeks of high-fat diet (HFD) or normal diet (ND) with densitometric analysis of 5 animals per group displayed as mean and SEM. Statistics: (b and d) paired t test, e one-way ANOVA with Tukey correction *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001