FIG. 5.
(A) LPS up-regulates caveolin-1 but not TLR4. TG-elicited macrophages were prepared from C3HeB/FeJ mice. Cells (106) were incubated with 1.0 pg/ml, 100 pg/ml, and 10 ng/ml LPS for 6 h. Equal volumes of total cell lysates were subjected to SDS-PAGE and analyzed by immunoblotting using rabbit anti-caveolin-1 antibody and rabbit anti-TLR4 antibody for detecting caveolin-1 and TLR4, respectively. (B) Sucrose gradient fractionation of caveolin-1 and TLR4. TG-elicited macrophages were stimulated with 1.0 ng/ml of LPS. Cells were then lysed, applied to a sucrose density gradient, and centrifuged as described in Materials and Methods. Fractions were subjected to SDS-PAGE. The presence of caveolin-1 and TLR4 was detected with rabbit anti-caveolin-1 and rabbit anti-TLR4 antibodies.