Fig. 3

Myofibroblast precursors are identified in the CD271+ subset. RNAseq analysis was performed on paired progenitor subsets isolated from SCAT of five independent obese donors. a Sparse partial least-square discrimant analysis plot of the two principal components. The 95% confidence ellipses are shown to strengthen the cell population clustering (left) and heatmap (right) of the top 400 most differentially expressed genes defining a progenitor subset-specific profile. Color code is set up according to gene expression (high expression in red, low in blue). b Gene ontology pathways determined on the 400 progenitor subset-specific genes. c Venn diagrams of differentially expressed genes (fold change ≥ 2 from a raw P-value) in −/− cells vs. −/CD271+ and MSCA1+ (left), in −/CD271+ cells vs. −/− and MSCA1+ (middle) and in MSCA1+ cells vs. −/− and −/CD271+ (right), highlighting some genes of interest. d Top ten superpathways determined from SC −/− subset-specific transcripts. e GLI1 and CD36 gene expression in native progenitor subsets determined by RT-qPCR analyses. The results are means ± s.e.m. of experiments performed on n = 6 (CD36) and n = 11 (GLI1) independent donors, one-way ANOVA followed by Tukey’s multiple comparison test, *P < 0.05. f Sorted SC progenitor subsets were cultured in the presence or absence of 5 ng/mL of TGFβ1 for 4 days, and immunostaining approaches were performed using phalloidin (F-ACTIN, green), αSMA antibody (red), and DAPI (blue), or cultured under adipogenic condition and stained with Bodipy (green) and DAPI (blue). Representative fluorescence microscopy images are shown, scale bar: 50 µm. g Gene expression levels in SC immunoselected progenitor subsets (−/−, −/CD271+, and MSCA1+ cells) treated with TGFβ1 were determined by RT-qPCR analyses. The results are expressed as log2 ratio of paired MSCA1+ or −/CD271+ cells to −/− cells mRNA levels and are means ± s.e.m. of experiments performed on n = 3 (COL6A2) to n = 7 independent donors, one-way ANOVA followed by Tukey’s multiple comparison test, *P < 0.05, **P < 0.01. h Quantification of lipid accumulation in SC progenitor subsets at day 10, the results are means ± s.e.m. of experiments performed on n = 4 independent donors. One-way ANOVA followed by Dunnett’s multiple comparison test, **P < 0.01