FIGURE 4.

Clearance and removal of Tollip from late endosomes/lysosomes by super-low-dose LPS. A, WT macrophages were treated with PBS or 50 pg/ml LPS for 1 h. Total cell lysates were harvested, and equal amounts of protein lysates were resolved by SDS-PAGE. The levels of Tollip and control GAPDH were visualized by Western blotting. WT macrophages were treated with 50 pg/ml LPS for either 1 h (B) or 24 h (C). Cells were stained with goat anti-mouse Tollip antibody, followed by Alexa Fluor 488-labeled rabbit anti-goat IgG together with either Cy3-conjugated LAMP1-specific antibody (B) or MitoTracker Red (C). The intracellular distribution of Tollip in late endosomes/lysosomes or mitochondrial compartments was visualized by confocal fluorescence microscopy.