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.
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).