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Fig. 8. Addition of exogenous PA activates cellular PLD. (A) Time course. 375 µM C₈-PA was added to MDCK cells and the cellular PLD activities assayed as described in Materials and Methods at the indicated times after addition. The values are means of duplicate determinations and the error bars are the ranges of the values. (B) Effects of various agents on PLD activation by C₈-PA. Cells were stimulated with 375 µM C₈-PA after 4 hours pre-incubation with 50 µM brefeldin A, or 15 minutes preincubation with 10 µM UO126 or 10 µM tyrphostin AG 1478. PMA was used at 0.1 µM in the presence or absence of 10 µM C₈-PA. LysoC₆-PA was used at 10 µM and 1-oleoyl-glycero-3-phosphate (lyso phosphatidic acid; LPA) complexed with lipid-free bovine serum albumin was used at 2 µM. This experiment was performed in the absence of serum. (C) Lack of increase of PtdIns(4,5)P₂ levels by C₈-PA was assayed at the indicated times after addition of 500 µM C₈-PA to MDCK cells. `5'kin' cells were infected with adenovirus coding for phosphatidylinositol 4-phosphate 5'-kinase as a positive control.

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