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. 2013 Jan 31;8(1):e54931. doi: 10.1371/journal.pone.0054931

Figure 2. p140Cap tyrosine phosphorylation depends mainly on two tyrosine residues. A.

Figure 2

Schematic representation of p140Cap structure and localization of FYELE, EPLYA, EGLYA, and ADPYG sequences into the tyrosine rich region. These four tyrosine residues have been mutated to phenylalanine. B. cDNAs encoding GFP, GFP-p140Cap full length (p140 WT) and its single (p140 EPLY/FA, p140 EGLY/FA), double (p140 EPLY/FA, EGLY/FA; p140 FY/FELE, ADPY/FG) and triple (p140 EPLY/FA, EGLY/FA, ADPY/FG) mutants were used to transfect HEK-293 cells. 48 hours after transfection cells were treated for 5 minutes with 100 micromolar pervanadate solution. Cell extracts were immunoprecipitated with a specific antibody to p140Cap and immunocomplexes were analysed by western blotting using monoclonal antibodies to phosphotyrosine (PY99), p140Cap and Tubulin respectively. The results are representative of six independent experiments.