Figure 3.

HGF promotes phosphorylation of β-catenin at Y142. A, Representative Western blots of crude neocortical synaptosomes following stimulation with HGF stimulation for 0, 5, 10, and 20 min. This experiment was repeated three times using independent synaptosomal preparations. The level of β-catenin phosphorylated at Y142 (p142-β-cat) increased in the presence of HGF, peaking at 5 min. Note the expected increase, followed by a time-dependent decrease, in phospho-MET (pMET) levels in response to HGF. Total levels of β-catenin (β-cat) and MET are unchanged. B, Representative confocal microscopy images of primary neocortical neurons at 14 DIV following stimulation for 5 min with PBS or HGF (25 ng/ml). This experiment was repeated in two independent culturing sessions. Note the increase in immunostaining of p142- β-catenin in the presence of HGF. Scale bar, 20 μm. C, Quantitative analysis of the p142- β-catenin clusters. Error bars represent the SEM; N = 26 cells from two independent cultures in each group. *p < 0.05 (HGF vs PBS).