Fig. 3

Impact of ML141 on hepatic, adipogenic, and neurogenic differentiation of young and aged-derived hADSCs. hADSCs were isolated from young or aged subjects and subjected to hepatocyte/adipocyte/neural differentiation as indicated in the Methods for 28 days. Cells were collected at day 0 (MSCs) at the moment of induction of the differentiation, and days 14 and 28 of the differentiation. Cells were incubated with or without 10 μM ML141 following two protocols of treatment: from day −2 to day 14 of the differentiation (d-2/14) and from day 14 to day 28 of the differentiation (d14/28). The effect of ML141 was evaluated at day 28 of the differentiation. a Representative images of morphological cell changes from: undifferentiated MSCs at D0, hepatoblast-like cells (HBLCs) or immature adipocyte-like cells (IALCs) or neurospheres (NSPs) at D14, hepatocyte-like cells (HLCs) or mature adipocyte-like cells (MALCs) or neural-like cells (NLCs) at D28. Cells from adipocyte differentiation were stained with oil-red O. b Hepatic/adipogenic/neurogenic marker expression: cell lysates (80–150 μg of protein) were separated by SDS-PAGE and immunoblotted with antibodies raised against cytokeratin (CK)-18, albumin (ALB), alpha fetoprotein (AFP), peroxisome proliferator activated receptor (PPAR)γ, FABP4, Pref-1, NeuN, O4, GFAP, and GAPDH. Protein expression profiling was determined during differentiation at D0/14/28; representative blots are shown