FIGURE 7.

bLF is endocytosed by LRP1. A, ST2 cells were incubated with the indicated concentration of bLF or PBS for 4 h. Whole cell lysates were immunoblotted with the anti-bLF antibody (left panel). ST2 cells were pretreated with 10 μg/ml bLF or PBS for 1, 2, or 4 h (middle panel). The inhibitory effects of bLF on the NFκB pathway at different incubation periods were examined by immunoblotting with an anti-IkBa antibody (right panel). B, ST2 cells were transfected with control, CD14, nucleolin (Ncl), or LRP1 siRNA. Thereafter, cells were incubated with 10 μg/ml bLF for 4 h. Whole cell lysates were immunoblotted with the anti-bLF antibody. Immunoblotting of β-actin was performed to control for protein loading. The experiments were performed at least twice, with similar results.