FIGURE 5.

Tyrosine kinase Src in vitro phosphorylation of OGT. OGT overexpressed from insect cells was incubated with ±Src in Src kinase buffer containing [³²P]ATP for 15 min. Reactions were stopped with modified Laemmli buffer, electrophoresed on 10% SDS-PAGE, and either stained with Coomassie Blue G250 and underwent an audioradiogram or was transferred to polyvinylidene difluoride for Western blot analysis with OGT (AL-28). Gel bands of OGT incubated with ±Src were cut out of SDS-PAGE, and [³²P]ATP was counted via a scintillation counter. Data are representative of experiments done in triplicate.