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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 (
) 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.