Figure 3.
Effects of 12(S)-HETE on Ang II binding. RMC were treated with 12(S)-HETE (10−7 M) for 24 h and (A) saturation binding assays with increasing amounts of [125I]-Ang II were performed on intact cells cultured in 24-well dishes. Unlabeled-Ang II (10−6 M) was used to define nonspecific binding. Samples were incubated for 90 min at 22°C, followed by 3 washes with PBS. Cells were lysed with NaOH and counted in a gamma counter. (B) Specific binding affinities were calculated from Scatchard plots using Prism software. (C) Ang II binding examined in cells treated with or without 12(S)-HETE for 2, 6, or 24 h. Bar graph represents mean ± SEM of three independent experiments (*P < 0.05 versus control).