Fig. 7

VS progenitors exhibit distinct differentiation potentials. RNAseq analysis was performed on paired progenitor subsets isolated from VS and SCAT of five 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 Venn diagrams of differentially expressed genes (fold change ≥ 2 from a raw P-value) between VS progenitor subsets: −/− cells vs. −/CD271+, and MSCA1+ (left), −/CD271+ cells vs. −/−, and MSCA1+ (middle) and MSCA1+ cells vs. −/− and −/CD271+ (right), highlighting some genes of interest. c Gene expression levels of BMP7 and CD24 in progenitor subsets isolated from SC and VSAT determined from RNAseq analyses. The results are expressed as log2 gene expression, n = 5 independent donors. Two-way ANOVA followed by Sidak’s multiple comparison test, *P < 0.05, **P < 0.01, ***P < 0.001. d Sorted VS 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 bars: 50 µm. e Gene expression levels of COL1A1, COL3A1, COL6A2, ELN, and GLI1 in VSAT −/− and –/CD271+ progenitor subsets treated or not with TGFβ1 were determined by RT-qPCR analyses. The results are expressed as means ± s.e.m. of experiments performed on n = 3 independent donors. One-way ANOVA followed by Sidak’s multiple comparison test, *P < 0.05, **P < 0.01, ***P < 0.001. f Quantification of lipid accumulation in VS progenitor subsets at day 10, results are means ± s.e.m. of experiments performed on n = 4 independent donors. One-way ANOVA followed by Tukey’s multiple comparison test, *P < 0.05