Novel Binding Partners and Differentially Regulated Phosphorylation Sites Clarify Eps8 as a Multi-Functional Adaptor
Figure 3
Differential regulation of tyrosine phosphorylation on Eps8.
Heavy, medium and light SILAC labelled HEK 293T cells were treated with either 25 nM dasatinib, 20 µM SU5402, or no inhibitor, prior to FGF2 stimulation (20 ng/ml; 15 min). Myc-Eps8 was immunoprecipitated and the resulting sample was run on an SDS-PAGE gel and, following in-gel trypsin digestion and phophopeptide enrichment, analysed by mass spectrometry. Each graph represents specific residues on Eps8 as indicated. Each data point represents a single peptide identification. P values were calculated by an unpaired t-test (0.01–0.05 = *; 0.001–0.01 = **; <0.001 = ***). +, the median of the ratios is<the cut-off value of 0.57 and is deemed significantly changed (see Method S1). A) Experiment was carried out in the absence of sodium pervanadate. B) Experiment was carried out in the presence of 2 mM sodium pervanvadate.
