FIGURE 7.

In vitro phosphorylation of UGT2B15 expressed in SYF^−/− cells by PKCα and Src kinase. Solubilized non-transfected and 2B15-transfected SYF^−/− fibroblast cells were prepared for kinase reaction. [γ-³³P]ATP was included all reactions, and kinase buffer, PKCα, Src kinase, or both kinases were added as described under “Experimental Procedures.” Reactions proceeded for 30 min at 30 °C. Experiments were carried out three times. Samples were purified by His affinity chromatography and subjected to resolution in a 4–15% gradient SDS-PAGE system. Gels were dried and exposed for photographs. H, His.