Figure 3

Cav-1 redistribution in HAECs co-cultured with HIV-infected cells during HDL mediated reverse cholesterol transport. HAECs were cultured in regular ECM (basal medium) (a), or serum free medium containing 1% fatty acid free BSA and cholesterol (40 µg/ml) for 36 h, and then further incubated in serum free medium without HDL (b) or with HDL (c) for 1–2 h. HAECs were then subjected to sucrose density gradient fractionation. Cav-1 distribution from discontinuous sucrose density gradient was examined by immunoblots. d HAECs were co-cultured with HUT-78HIV cells for 5 days, and HAECs were placed in regular ECM medium (cocul/HUT78HIV basal). After 5 days co-culturing with HUT-78HIV, HAECs were placed in serum free medium in the presence of cholesterol (40 µg/ml) and 1% fatty acid free BSA, and then followed incubation in medium e without HDL or f with HDL for 1–2 h. Using Na₂CO₃ based sucrose density gradient fractionation, 12 fractions were obtained and examined by Western blotting for Cav-1 distribution. Representative data are shown from three independent experiments. The ratio of Cav-1 distribution from mock or HUT-78HIV co-cultured HAECs in Cav-1 enriched membrane fractions 4–5 and non-caveolar fractions 6–8 and 9–12 based on three independent experiments are shown at the bottom. Dash lines refer to caveolae enriched membrane fractions 4 and 5.