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. 2020 May 7;27(10):2921–2941. doi: 10.1038/s41418-020-0551-y

Fig. 3. β-catenin degradation marks FAPs undergoing adipogenesis.

Fig. 3

a Contour maps obtained by projecting, onto a two-dimensional plane, single-cell multiparametric data using the viSNE algorithm. Data from three independent FAP preparations, in the presence or absence of ADM, are shown. b Dot plot representations of the viSNE maps showing β-catenin expression (color coded, blue = low, red = high) across the reported conditions for each repeat. c Representative western blot showing the protein levels of both PPARγ isoforms, non-phospho (active) β-catenin and Vinculin in FAPs unstimulated or induced to differentiate into adipocytes upon ADM incubation (n = 3). d Bar graphs representing the densitometric values of PPARγ1, PPARγ2 in both conditions. e Bar graph showing the protein levels of non-phospho (active) β-catenin in both conditions. Protein levels were normalized to Vinculin. The statistical significance was estimated by Student's t test. f Relative expression of Ctnn1b in FAPs unstimulated or induced to differentiate into adipocytes upon ADM exposure (n = 3). g Western blot showing the protein levels of PPARγ, non-phospho (active) β-catenin and Vinculin in FAPs undergoing adipogenesis in the absence or presence of 20 μM MG132 (n = 3). (h) Bar graphs representing the densitometric values of PPARγ and non-phospho (active) β-catenin in both conditions. Protein levels were normalized to Vinculin. The statistical significance was estimated by Student's t test. i Western blot showing the protein levels of PPARγ, non-phospho (active) β-catenin and Vinculin in FAPs unstimulated or incubated with ADM and ADM supplemented with 20 nM LY2090314 (n = 3). j Bar graphs representing the densitometric values of PPARγ and non-phospho (active) β-catenin in all conditions. Protein levels were normalized to Vinculin. The statistical significance was estimated by One-way ANOVA. k Representative immunofluorescence (left) (20× magnification; scale bar, 100 μm) of TAs of young (1.5-month-old, n = 6) and old (18-month-old, n = 7) mdx mice. Infiltrating adipocytes (arrow heads) were revealed using antibodies against perilipin-1 while fibers were stained using phalloidin-488. Bar plot (right) reporting the fraction of perilipin positive area in young and old mdx TAs. Statistical significance was estimated by Student's t test. l Immunoblot showing the protein level of non-phospho (active) β-catenin, pGSK3 (Ser9) and Vinculin (n = 5) in FAPs from young (1.5-month-old) and old (18-month-old) mdx mice. m Bar plots showing the densitometric analysis for non-phospho (active) β-catenin and pGSK3 (Ser9) normalized over Vinculin, respectively. n Quantitative PCR (qPCR) for Ctnnb1 from FAPs purified from young (1.5-month-old) and old (18-month-old) mdx mice (n = 3). Statistical significance was estimated by Student's t test. All data are represented as mean ± SEM and the statistical significance is defined as *p < 0.05; **p < 0.01; ***p < 0.001.