Lipopolysaccharide Stimulates p62-Dependent Autophagy-Like Aggregate Clearance in Hepatocytes
Figure 1
LPS induces TLR4-mediated increases in autophagy in liver and hepatocytes. (a) Immunoblot for LC3II in liver lysates from WT (C57BL/6) mice given no LPS (0 h) or LPS 5 mg/kg ip for 6 or 24 h. Each lane represents liver from one mouse. (b) Immunoblot of LC3II in whole cell lysates from primary mouse hepatocytes treated with LPS (100 ng/mL) for up to 24 h (/gp). (c) LC3-puncta formation with quantification (below) in primary mouse hepatocytes transfected with GFP-LC3 prior to stimulation with LPS (100 ng/mL) for up to 24 h. (d) LC3-puncta formation with quantification (below) in primary mouse hepatocytes pretreated with or without bafilomycin for 1 h plus transfection of GFP-LC3 prior to stimulation with LPS for 16 h. (e) Immunoblot for LC3II in hepatocytes from WT and TLR4ko mice treated for up to 24 h with 100 ng/mL LPS. (f) LC3-puncta formation with quantification (right hand graph) in WT and TLR4ko primary hepatocytes transfected with GFP-LC3 without LPS (0 h or no treatment) and stimulated with LPS (100 ng/mL) for 16 h. versus WT LPS 16 h; data shown as mean ± SEM; images representative of at least 3 separate experiments. Confocal immunofluorescence ×60 magnification, quantification using MetaMorph of GFP-positive puncta ≥100 cells normalized to numbers of nuclei.
