Fig 2: The miR-27a-3p target LPL does not alter adipogenesis. To assess if miR-27a-3p regulated its predicted target, lipoprotein lipase (LPL), SGBS preadipocytes were transfected 48 h prior to adipogenic induction with miRNA mimics or non-target control (NT, 20 nM), and additionally, a dual-luciferase reporter assay was performed in HEK293 cells. (A) mRNA level of LPL as quantified by qPCR normalized to HPRT at indicated time points of adipogenic differentiation. (B) One representative Western blot from three independently performed experiments for LPL using tubulin as loading control. (C) Illustration of the predicted binding site for miR-27a-3p in the 3' UTR in the human LPL mRNA. (D) Results of the dual-luciferase reporter assay performed in HEK293 cells. Luminescence from firefly/Renilla luciferase activity was determined as indicated. (E–G) SGBS preadipocytes were transfected 48 h prior to adipogenic induction with either control non-target (Ctrl) or an siRNA pool targeting human LPL (si-LPL, 20 nM). RNA and protein samples were collected on day 14 of the differentiation process. (E) mRNA expression of LPL and adipogenic markers as quantified by qPCR normalized to HPRT after LPL knockdown. (F) One representative Western blot out of three independent experiments after LPL knockdown in SGBS cells. (G) Densitometric analysis of all Western blots on day 14 of adipogenesis normalized to tubulin. Statistics: results are displayed as means and SEMs of 5 (A) and 3 (B,D–G) independent experiments. Two-way ANOVA with Bonferroni correction (A); t-tests (D,E,G) with respect to NT/Ctrl for the same time point; * p < 0.05; *** p < 0.001. HPRT: hypoxanthine-guanine phosphoribosyltransferase; FASN: fatty-acid synthase; PLIN1: perilipin; PPAR?: peroxisome proliferator-activated receptor ?; ADIPOQ: adiponectin; 3p: miR-27a-3p.