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Fig. 2. Cholesterol efflux from J774-A1 macrophages in the presence of native and tyrosylated HDL3 (A) or total HDL (B). J774-A1 cells were [3H]cholesterol-loaded with [3H]cholesteryl oleate-labeled AcLDL (50 µg apo B/ml, 7.6x105 DPM/ml) as described in Materials and Methods. After an 18 hour equilibration period, the cells were incubated for 24 hours at 37°C with the indicated final concentrations of native (•) or tyrosylated ({square}) HDL3 (A) or total HDL (B). After extensive washing with PBS, cellular and medium lipids were extracted, separated by TLC and counted to determine the [3H]cholesteryl ester and the [3H]free cholesterol fractions. Cholesterol efflux was calculated as the amount of [3H]cholesterol (free and esterified) released to the medium relative to total label in each well. Each value is the mean±s.e.m. of three determinations. *P<0.03, **P<0.02, ***P<0.01.





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