Figure 6

Cav-1 localization in Nef treated HAECs. a Cav-1 distribution was examined in HAECs cultured in regular ECM basal medium using sucrose density gradient subcellular fractionation and immunoblots. b Cav-1 distribution in HAECs incubated in medium containing 40 µg/ml cholesterol and 1% fatty acid free BSA. c Cav-1 distribution in fractions was measured in cells pre-incubated with cholesterol for 36 h followed by incubation in serum free medium containing HDL for 2 h. d Cav-1 localization in sucrose density fractions was examined after HAECs were incubated in regular ECM medium (basal) in presence of HIV Nef for 3 days. e HAECs were treated with Nef for 3 days, then incubated in medium containing 40 µg/ml cholesterol and 1% fatty acid free BSA for 36 h, followed by a 2 h incubation in serum free medium without or f with HDL. Ratio of Cav-1 distribution from mock or Nef treated HAECs in Cav-1 enriched membrane fractions 4–5 and non-caveolar fractions 6–8 and 9–12 is shown at the bottom. Dash lines refer to caveolae enriched membrane fractions 4 and 5. Representative data are shown from three independent experiments.