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

Figure 3

Evaluation of KGF effect on ASC proliferation. (a) Proliferation ability of ASCs treated or not with 20 ng/ml KGF for 1–5 days was determined by MTT assay. Error bars represent standard deviations from three independent experiments. and . (b) Phosphorylation of ERK was assessed by Western blot analysis with a phospho-specific ERK monoclonal antibody (pERK) on ASC whole cell lysates, treated or not with 20 ng/ml KGF for 5 and 30 min. Levels of total ERK were assessed by blotting with an ERK2-specific antibody and served as loading control. (c) Phosphorylation of p38 was assessed by Western blot analysis with a phospho-specific p38 monoclonal antibody (pp38) on ASC whole cell lysates, treated or not with 20 ng/ml KGF for 5 and 30 min. Levels of total p38 were assessed by blotting with a p38-specific antibody and served as loading control. (d) Phosphorylation of ERK was assessed by Western blot analysis with a phospho-specific ERK monoclonal antibody (pERK) on ASC whole cell lysates, treated or not with 20 ng/ml KGF for 5 min, in the presence or not of the p38 inhibitor SB202190 or of the ERK inhibitor U0126. Levels of total ERK were assessed by blotting with an ERK2-specific antibody and served as loading control. (e) Phosphorylation of p38 was assessed by Western blot analysis with a phospho-specific p38 monoclonal antibody (pp38) on ASC whole cell lysates, treated or not with 20 ng/ml KGF for 30 min, in the presence or not of the p38 inhibitor SB202190 or of the ERK inhibitor U0126. Levels of total p38 were assessed by blotting with a p38-specific antibody and served as loading control. (b–e) The intensity of the bands was evaluated by densitometric analysis; the values from at least three independent experiments were normalized and reported as fold increase with respect to the untreated sample. (f) Proliferation ability of ASCs treated or not with 20 ng/ml KGF in the presence or not of the p38 inhibitor SB202190 or of the ERK inhibitor U0126 for 3 days was determined by MTT assay. Error bars represent standard deviations from three independent experiments. and .

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