Figure 5.

In vitro phosphorylation of Par-4 wt and different Par-4 phospho-mutants by Dlk. Purified recombinant Par-4 wt (2 μg, lane 1), Par-4 S154A (lane 2), Par-4 T155A (lane 3), and Par-4 S154A/T155A (lane 4) were phosphorylated by recombinant Dlk in the presence of [γ-³²P]ATP, analyzed on 10% SDS-PAGE, and visualized by autoradiography (top). Equal protein loading of all Par-4 constructs was demonstrated by Coomassie staining of the same gel (bottom). For better depiction of differences in signal intensities a shorter exposure time was chosen for autoradiography than in Figure 3. Hence autophosphorylation of Dlk is not visible.