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. 2021 Jan 20;23:1035–1052. doi: 10.1016/j.omtn.2021.01.014

Figure 7.

Figure 7

Effect of miR-223-3p overexpression on glucose metabolism in adipocytes at a late stage of differentiation

(A–D) mRNA expression of Glut4 (A); representative blot and quantification of GLUT4 (B); representative blots and quantification of IRS-1, pIRS-1(Tyr608), Akt, pAkt(Ser473), AS-160, pAS-160(Ser666), and their respective ratios (C); and representative blot and quantification of ARF6 (D) in 3T3-L1 cells transfected with negative miRNA-control (control) or miR-223-3p mimic (mimic) at day 6 of differentiation. (E and F) Representative confocal micrographs and quantification of GLUT4 translocation (E) and glucose uptake (F) of cells transfected with negative miRNA-control (control) or miR-223-3p mimic (mimic) at day 6 of differentiation, stimulated or not with insulin (100 nM, 20 min). Cells were co-transfected with cMyc-GLUT4-EGFP expression plasmid and immunostained with anti-cMyc antibody, showing GLUT4t in green and GLUT4m in red. Nuclei were stained with DAPI (blue). Scale bars, 10 μm. Values are given as mean ± SEM (n = 6), ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 versus negative miRNA control; ##p < 0.01 and ###p < 0.001 versus noninsulin-stimulated cells. See also Figures S4D and S4E.