MSDC-0602K decreases adipocyte differentiation potential of BM-MSCs compared to pioglitazone in obese mice. (A) Representative picture of Oil red O (ORO) stained lipid droplets in AD differentiated mBM-MSCs (scale bar 150 μm; 20x magnification). (B–C) Gene expression profile of mBM-MSCs differentiated towards adipocytes in D10. (B) Gene expression of adipogenic genes (Pparγ2, Cebpa, Adipoq, Cd36, Fsp27, Insr). Data are presented as mean fold change (F.C.) of gene expression normalized to mBM-MSCs from ND group ± SEM (n = 2 from pooled samples); one-way ANOVA, Tukey's multiple comparison test, a: ND vs other groups; b: HFD vs other groups, c: HFD + P vs other groups. (C) Representative pictures of Alizarin Red (AZR) staining for calcified matrix mineralization of OB differentiated mBM-MSCs (scale bar 150 μm; 20x magnification) and (D) quantification of eluted AZR staining of mineralized matrix in OB differentiated mBM-MSCs in D10. Data are presented as mean fold change (F.C.) of A500 in OB differentiated cells (OB) normalized to mBM-MSCs from ND group ± SEM (n = 2–10); one-way ANOVA, Tukey's multiple comparison test, a: ND vs other groups; b: HFD vs other groups and unpaired t-test ∗∗p < 0.01: HFD + P vs HFD + M. (n = 2–10). (E) Measurement of alkaline phosphatase (ALP) activity normalized to cell viability in OB differentiated mBM-MSCs in D7. Data are presented as mean fold change of ALP activity in OB differentiated cells (OB) from each experimental group ± SEM (n = 7–14); one-way ANOVA, Dunett's test comparing a: ND vs other groups and unpaired t-test ∗∗p < 0.01: HFD + P vs HFD + M. (F) Gene expression profile of osteoblastic markers (Alpl, Oc, Col1a1 and Bmp2) and (G) Dlx5, Msx2, Ctnnb1, Lrp5, in OB differentiated mBM-MSCs in D10. Data are presented as mean fold change (F.C.) of gene expression normalized to mBM-MSCs from ND group ± SEM (n = 2 from pooled samples); one-way ANOVA, Tukey's multiple comparison test, a: ND vs other groups, b: HFD vs other groups, c: HFD + P vs other groups, d: HFD + M vs other groups.