In vitro kinase assays were performed as described previously9 (link). Wild-type and T579A His-tagged FE65 PTB2 (residues 531 to 666) recombinant proteins were expressed and purified from E. coli as substrates. HA-GSKβ was isolated from transfected cell lysate by immunoprecipitation using anti-HA 12CA5 antibody (Roche).
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