Figure 2.

AngII and exogenous bacterial PLD increase [³H]myristate-labeled DAG that persists after agent removal. Primary cultures of bovine adrenal glomerulosa cells in serum-free medium for 20–24 h were labeled for 2 h in KRB⁺ containing 5 μCi/ml [³H]myristic acid. The KRB⁺ was removed and replaced with KRB⁺ containing no additions (control, Con) or with 10 nm AngII (A) or 0.1 IU/ml PLD (B) for 30 min as shown. Adrenal glomerulosa cells were then washed three times with KRB⁺ before incubation for an additional 30 min with KRB⁺ alone or KRB⁺ containing 10 nm AngII (A) or 0.1 IU/ml PLD (B) as indicated. Radiolabeled DAG was then extracted into chloroform/methanol, separated by thin-layer chromatography, and quantified as described in Materials and Methods. Values represent the means ± sem of 10 samples from five separate experiments. ***, P < 0.001; *, P < 0.05 vs. control (without agent in both the first and second 30-min time periods).