Fig. 9

Induction of Hep-Dif in Cdc42 knockdown aged human adipose-derived mesenchymal stem cells (hADSCs). hADSCs derived from aged subjects were transfected with an Cdc42-targeting siRNA (siCdc42) at day –2 (D–2) and then differentiated at day 0 (D0) with 1% FBS. Cells were incubated with or without NSC5476 (NSC, 5 μM) 24 h before launching the differentiation and maintained along the Hep-Dif. Cdc42 mRNA and Cdc42-GTP protein levels as well as key factors were assessed during hepatocyte differentiation of Cdc42-knockdown cells from D<0 to D28: secreted albumin protein, released exosomes, miR122 mRNA, hepatocyte nuclear factor (HNF)4 mRNA, and E-cadherin mRNA. Results are normalized as previously to the expression of the housekeeping gene and are the mean ± SEM of three independent experiments and are expressed as the percentage or fold variation of control relative to day D–2 (considered as D<0). *P < 0.05, **P < 0.01, ***P < 0.005, D0/D14/D28 vs D < 0; ^§P < 0.05, ^§§P < 0.01, NSC vs control