Figure 5.

In Vivo Labeling with ³²Pi Indicates a Rapid Rate of [³²P]PtdInsP₂ Biosynthesis in the Hs PIPK1α Lines.

All cells were preequilibrated in conditioned medium for 30 min; ³²Pi was added, cells were harvested, and lipids were extracted at the time points indicated. The lipids were separated by TLC, and ³²P-labeled lipids were quantified with a Bioscan imaging scanner.

(A) Representative autoradiogram of the TLC plate.

(B) to (D) ³²P recovered phospholipids (PtdInsP₂, PtdInsP, and PtdOH, respectively) over the time course (wild type, open diamonds; Hs PIPKIα lines, closed circles). The data are reported as percentage of total cpm recovered per lane. Each point is the average ± sd of duplicates from three independent experiments.