Research Paper Advance Articles

Figure 4. miR-145-5p and miR-27b-3p inhibitors enhance adipogenic differentiation in HGPS cells. HGPS cell strains (HGADFN003, HGADFN127, and HGADFN164) were transfected with 10 nM hsa-miR-145-5p and/or has-miR-27b-3p inhibitors on days 0, 3, 6, and 9 during SKP-to-adipocyte differentiation. The treatments groups include 10 nM negative control inhibitor (NC (In)), 10 nM hsa-miR-145-5p inhibitor (145 (In)), 10 nM hsa-miR-27b-3p inhibitor (27b (In)) and a combination of 10 nM hsa-miR-145-5p inhibitor and 10 nM hsa-miR-27b-3p inhibitor (Combi (In)); qPCR analysis was performed for miR-145-5p (A), and miR-27b-3p (B), both normalized to U6, as well as for IRS1 (C), KLF4 (D), KLF5 (E), PHB (F), PPARγ (G) and LPL (H) all normalized to GAPDH. (I) Bodipy staining (green) of lipid droplets on day 12 of differentiation with representative images (40x magnification; scale bar: 20 μm). DAPI was used as a counterstain. (J) Quantification of the percentage of Bodipy-positive area normalized to the total number of DAPI area. (K) Quantification of the percentage of Bodipy-positive cells normalized to the total number of DAPI-positive cells. (A–H) Values are presented as mean ± SD (n=3); p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; two-way ANOVA with Tukey’s multiple comparison test. (J, K) Values are presented as the mean ± SD (n=3); p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; Ordinary one-way ANOVA with Dunnett’s multiple comparison test.