FIG. 7.

Effect of exogenous kinases on the PLD activity of phosphorylation-deficient mutants. PLD2-Y296F (A, D, and G), PLD2-Y415F (B, E, and H), and PLD2-Y511F (C, F, and I) PLD2 mutants were transfected into COS-7 (A to C), MTLn3 (D to F), or MCF-7 (G to I) cells. After 36 h of expression, lysates were used for in vitro kinase reactions with EGFR, JAK3, or Src. After this, PLD2 was immunoprecipitated with α-myc antibodies, and samples were split into two 50-μl aliquots for duplicate determinations in the PLD2 assay. The results represent means ± the SEM of four independent experiments in duplicate. *, differences between means (no-kinase versus kinase treatments) that were statistically significant (P < 0.05) as determined by ANOVA.