Figure 2.
Angiotensin II promotes p47phox and p67phox mobilization to the cell membrane in human neutrophils. Neutrophils were suspended in KR-HEPES buffer at 107 cells/mL and were incubated with 100 nM angiotensin II (Ang) for short (until 10 minutes) (A) or longer (until 120 minutes) durations (B), or the cells were previously incubated in the absence or presence of PD098059 (50 μM) for 30 minutes and Ang II was then added at the indicated concentrations for 15 minutes (C). Cells were lysed, and the plasma-membrane–enriched fraction was separated from the cytosolic fraction by ultracentrifugation at 100 000g. Membrane proteins (50 μg/lane) were subjected to SDS-PAGE, transferred to PVDF membranes, and probed with antisera against p47phox and p67phox. Each blot is representative of a set of 3 experiments that yielded similar results.