Research Paper Advance Articles

Figure 2. Impaired adipocyte differentiation in HGPS compared to control cells. Control cell strains: GM05565, GM05757, and GM01651; HGPS cell strains: HGADFN003, HGADFN127, and HGADFN164; (A) Schematic outline of adipocyte differentiation protocol; (B) Bodipy staining of lipid vesicles (green) in control and HGPS adipocytes on day 12 (40x magnification; scale bar 20 μm) with DAPI counterstaining. (C) Quantification of the Bodipy-positive area, normalized to the DAPI signal area. (D) Quantification of the percentage of Bodipy-positive cells normalized to DAPI positive cells. (E) Representative western blot of Lamin A/C including progerin, PPARγ and FABP4 at day 3, 6, 9 and 12 of adipogenesis (for uncropped and unprocessed images, see Supplementary Figure 3). Quantification of PPARγ (F) and FABP4 (G) protein level normalized to GAPDH. qPCR analysis of miR-145-5p (H) and miR-27b-3p (I) normalized to U6 and mRNA levels of C/EBPα (J), PPARγ (K), and FABP4 (L) normalized to GAPDH. (J–L) Values are presented as the mean ± SD (n=3); p > 0.05; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; (B, C) unpaired t-test; (E–I) two-way ANOVA with Sidak’s multiple comparisons test.