Fig. 3

Autophosphorylation of DYRK1A point mutants in vitro. Wild type and mutant DYRK1A constructs were expressed in vitro by coupled transcription-translation (see ``Materials and methods''). Samples were incubated for 90 min at 37 °C before tyrosine autophosphorylation of the reaction products was assessed by immunoblot analysis a. DYRK1A-L295F was included as a control because this mutation is known to reduce tyrosine autophosphorylation in this assay [9] For quantification, pY321 signals were normalized to the total amounts of recombinant DYRK1A (b) (means and s.d., n = 3). The differences in relative tyrosine autophosphorylation between wild type and mutant were tested for statistical significance by One sample t test (*p < 0.05)