Cholesterol replenishment in caveolae/lipid rafts reverses DHA22:6,n3 inhibition of TNF-α induced ICAM-1 expression. hRVE cells treated with 100 μM DHA22:6,n3 (black bars) for 12 hours followed by TNF-α stimulation for 12 hours. Cells treated with palmitic acid16:0 were used as a control (gray bars). Water-soluble cholesterol (25 μM cholesterol complexed with 250 μM MCD) was added to DHA22:6,n3-treated cells for 30 minutes before TNF-α stimulation (striped bar). ICAM-1 expression levels were assessed by immunoblot analyses. Equal amounts of protein were added to each lane, as confirmed by actin levels. Data are the mean ± SD of results in three experiments, *P < 0.05 compared with palmitic acid16:0 treated cells. #P < 0.05 compared with DHA22:6,n3-treated cells without cholesterol.