Figure 3.

miR-669a-5p promotes the differentiation of 3T3-L1 cells. 3T3-L1 preadipocytes were transfected with mimic control or mimic miR-669a-5p (100 nM) on day 0 and day 4 after differentiation, cells were harvested on day 8 for analysis.(a) RT-qPCR to analyse the expression of miR-669a-5p in 3T3-L1 cells transfected with mimic control or mimic miR-669a-5p during differentiation, normalized to U6 expression. n = 3 per group.(b-c) Lipid accumulation was assessed by Oil Red O staining, and the absorbance was measured at 510 nm wave length. A representative image of three independent experiments is shown in B.(d) Lipid accumulation was assessed by intracellular TG content. n = 3 per group(e) RT-qPCR to analyse the mRNA levels of adipogenesis markers Pparγ, Fabp4 and perilipin2 in differentiated 3T3-L1 adipocytes transfected with mimic control or mimic miR-669a-5p, normalized to β-actin expression. n = 3 per group.(f) Western blot to evaluate the protein levers of adipogenesis markers PPARγ, FABP4 and PERILIPLIN2 in differentiated 3T3-L1 adipocytes transfected with mimic control or mimic miR-669a-5p, β-ACTIN was used as a loading control. n = 3 per group.(g) Quantitative densitometry of the Western blots showed in F.Data are representative of at least three individual experiments. Results are represented as mean ± SEM. **p < 0.01 versus mimic control group. Scale bar indicates 200 μm in B.