Fig. 4: Mitochondrial signature and oxidative capacity of adipogenic compared to SWAT cells.
From: Adipogenic and SWAT cells separate from a common progenitor in human brown and white adipose depots
a, Predicted brown and white adipocyte content in pseudotime trajectories using BATLAS. Cells are grouped by each developmental branch and pseudotime decile. b, BATLAS genes overlapping with this work’s scRNA-seq dataset, expressed in branches across depot origins. c, Adipogenic cells contain lipid droplets as indicated by immunofluorescence staining of differentiating adipocytes for perilipin (green) combined with FISH RNAscope for DCN (magenta) and ADIPOQ (yellow). Nuclei in blue. Scale bar, 100 μm. d, Illustration demonstrating density gradient centrifugation to enrich adipogenic and SWAT cells from heterogeneous cultures. Created with BioRender.com. e, Seahorse extracellular flux analysis of enriched adipogenic and SWAT cells separated on day 10 of differentiation. Oxygen consumption rates (OCRs) were measured 24 h after plating in DM (5,000 cells per well) and normalized to cell count. N = 3 biologically independent cell samples, cultured separately in parallel for 24 h following density gradient centrifugation between SWAT and adipogenic cells. f, Calculations of the data visualized in e. Two-way analysis of variance (ANOVA) was used to assess differences between cell types and NE treatment in: basal mitochondrial respiration (effect of cell type P = 0.0006; effect of NE: P = 0.222), NE-stimulated mitochondrial respiration (effect of cell type P = 0.0003; effect of NE: P < 0.0001), stimulated ATP production (effect of cell type: P = 0.0012; effect of NE: P = 0.3310) and stimulated proton leak (effect of cell type: P < 0.0001; effect of NE: P = 0.0001). N = 3 biologically independent cell samples, cultured separately in parallel for 24 h following density gradient centrifugation between SWAT and adipogenic cells. Sidak’s post hoc test was used for comparisons between cell types, and significance values are shown in the graphs *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data are presented as the mean ± s.e.m. NE, noradrenaline; oligo, oligomycin; R, rotenone; A, antimycin; DM, differentiation medium 2.
