Fig. 2.
Dose response of PMA and mezerein for 5-LO product formation and 5-LO translocation in MM6 cells.
Intact MM6 cells (3 × 106 in 1 mL PGC buffer) were primed for 10 minutes at 37°C with the indicated concentrations of PMA (●) or mezerein (○). Then cells were stimulated with 5 μM A23187 (A) or A23187 and AA (5 μM and 40 μM, respectively) (B) for 10 minutes, and 5-LO product formation was determined by HPLC. Results are given as mean + SE; n = 3-5. (C) MM6 cells (1 × 107 in 1 mL PGC buffer) were primed with the indicated concentrations of PMA or mezerein for 10 minutes at 37°C before stimulation with 5 μM A23187 for 5 minutes. Cells were fractionated, and nonnuclear and nuclear fractions were analyzed for 5-LO by immunoblotting as described. Similar results were obtained in one additional experiment.