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Figure 5


Fig. 5. Y737 in β2-adaptin is a regulatory site for Src-mediated binding of β-arrestin in vitro. (A) GST-fusion proteins of β2-adaptin with single substitutions (Y737F, Y874F and Y926F) were either left unphosphorylated (–) or phosphorylated (+) with purified Src. The amounts of GST-β2-adaptin were assessed by Ponceau Red and Src-phosphorylated proteins were detected by western blot. (B) After removing Src from the reaction, the unphosphorylated or phosphorylated GST-proteins were incubated with increasing amounts of Flag-β-arrestin1. The amounts of GST-proteins were detected using an anti-GST antibody and β-arrestin associated with the GST proteins was determined by western blot using anti-Flag antibody. Data are representative of three to five independent experiments.





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