Figure 4.

DHEA stimulates endothelial proliferation independent of eNOS activity. BAECs were incubated with vehicle (C) or 5% FBS for 24 h or 1 nm DHEA for 15 min at 37 C for 24 h. L-NAME (5 mm) was added 30 min prior and also coincubated for 15 min with or without DHEA. After 24 h the cells were fixed and immunostained for Ki 67 proliferative antigen (as described in Materials and Methods). Data from three independent experiments with triplicate samples, along with representative fields at ×20 magnification, are shown. Data were expressed as means ± sem of Ki67-positive cells as a percentage of total cells (stained and unstained for Ki67) from at least 3000 total cells counted per condition. *, P < 0.05 vs. vehicle (C).