Eugene A. Podrez, Maria Febbraio, Nader Sheibani, David Schmitt, Roy L. Silverstein, David P. Hajjar, Peter A. Cohen, William A. Frazier, Henry F. Hoff, Stanley L. Hazen
Effect of lipid competitors on the binding of NO2-LDL to
CD36-transfected cells. [125I]LDL was
modified as described for the complete system in Figure 2a.
[125I]-NO2LDL (5
μg/mL) was then incubated with CD36-expressing 293 cells for 3 hours
at 4°C in the presence of (a) 20 μg
lipid/mL or (b) the indicated concentrations (μg
lipid/mL) of competitors. PAPC, PAPC(SnCl2), PLPC, and POPC
unilamellar vesicles were oxidized for 8 hours at 37°C as described
for the complete system in Figure 2 in the presence
(+NO2–, filled symbols) or
absence (–NO2–, open symbols) of
NO2–. Where indicated, BSA (0.2 mg
protein/mL final concentration) was also included during liposome preparation as
described in Methods (hatched bars). PAPC (SnCl2), hydroperoxide-free
PAPC generated by reduction of PAPC with SnCl2, and then reisolation
of PAPC under argon atmosphere before use were used as described in Methods.
Data represent the mean ± SD of triplicate determinations
(a) or means of triplicate determinations (b) of a
representative experiment performed 3 times. AP
< 0.001 for comparison versus control (no competitor).