Figure 5.

Downregulation of Cav-1 enhances the differentiation of hADSCs into dopaminergic-like neurons.

(A) Protein levels of Cav-1. hADSCs were transfected with Cav-1 siRNA. After siRNA transfection for 72 hours, the expression of Cav-1 was assessed by western blot assay. (B) Quantification of Cav-1 protein expression. (C) Expression and subcellular distribution of TH, Lmx1a and Nurr1 by immunofluorescence: The red fluorescence indicates cells that are positive for TH, Lmx1a and Nurr1. The blue fluorescence indicates nuclei stained with hoechst 33258. Scale bar: 50 μm. (D, E) Western blot assay and quantitative analysis of the expression of TH, Lmx1a and Nurr1. (F) Quantitative analysis of mRNA expression after dopaminergic neuronal induction of hADSCs. (G) Quantitative analysis of the release of dopamine by the differentiated cells at 29 days and by Cav-1 KD cells at 29 days. &P < 0.05, &&P < 0.01, vs. Cav-1 KD 29 day. Data are presented as the mean ± SD (Student’s t-test; n = 3). Average of three independent in vitro differentiation experiments was calculated. Cav-1: Caveolin-1; hADSCs: human adipose-derived stem cells; Lmx1a: LIM homeobox transcription factor 1 alpha; TH: tyrosine hydroxylase; KD: knock down.