Figure 4

RU-486 counteracts budesonide or dexamethasone inhibition of FAP adipogenic differentiation. (a) WT or mdx FAPs were isolated by the standard procedure and plated in fGM. After 24 hours, cells were treated with 0.1, 1 or 5 μM of budesonide or dexamethasone either with or without RU-486. After 6 days, cells were stained with ORO to evaluate adipocyte formation. (b) Immunofluorescence showing ORO staining (red) following differentiation of mdx FAPs. Nuclei are stained with Hoechst 33342 and are shown in grey. (c) Bar plot showing the fraction of ORO positive cells for mdx FAPs. (n = 3–4) ± SEM. (d) Immunofluorescence microphotographs showing WT FAPs stained with ORO to reveal adipocytes (red) and Hoechst 33342 to reveal nuclei (grey). (e) Bar graphs presenting the fraction of ORO positive cells for the experiment reported in d. The values are mean of three independent experiments ± SEM (n = 3) Statistical significance has been evaluated using one-way ANOVA (*p ≤ 0.05, **p ≤ 0.01, ns: not significant). Scale bar: 100 μm.