Neuropilin 1 Mediates Keratinocyte Growth Factor Signaling in Adipose-Derived Stem Cells: Potential Involvement in Adipogenesis

Figure 4

Evaluation of KGF effect on ASC adipogenic differentiation. (a) HPTLC analysis of the neutral-lipid cholesterol (CHOL), triglycerides (TGs), and cholesterol esters (CEs) in ASCs treated or not with 20 ng/ml KGF for 24 h. (b) The intensity of the bands was evaluated by densitometric analysis, normalized and reported in a graph as relative expression with respect to untreated cells. Error bars represent standard deviations (). (c) Representative images of ASCs cultured in adipogenic medium with or without KGF for 21 days and subjected to lipid staining with oil red O. Stained cells were then solubilized using isopropanol, and the extent of adipocyte differentiation was quantitated by determining the amount of extracted dye, as measured by the optimal absorbance at 490 nM, reported in the graph as relative expression with respect to untreated cells (). (d) Immunofluorescence analysis of the adipocyte-specific fatty acid binding protein 4 (FABP4, red) in ASCs at day 21 of adipogenic differentiation, treated or not with KGF during differentiation. Nuclei (blue) were visualized with 4,6-diamidino-2-phenylindole (DAPI). The percentage of FABP4-positive cells was determined by counting the number of FABP4-positive cells versus total number of cells in ten different areas randomly taken from three independent experiments. Error bars represent standard deviations (). (e) Relative PPARγ mRNA levels at day 21 of adipogenic differentiation in ASCs treated with KGF are shown as fold value of the level of PPARγ mRNA in untreated cells. Error bars represent standard deviations. (f) Relative FABP4 mRNA levels at day 21 of adipogenic differentiation in ASCs treated with KGF are shown as fold value of the level of FABP4 mRNA in untreated cells. Error bars represent standard deviations ().

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