Figure 3

Exposure to LPS diminished neutrophil chemotaxis and AMPK phosphorylation. (A) Representative images show neutrophil transmigration before and after treatment with LPS. Neutrophils were treated with LPS (0 or 1 µg/mL) for 90 min. The cells were then washed and chemotaxis examined over a 60 min period. In (B), neutrophils were incubated with LPS for 90 min followed by measurement of neutrophil chemotaxis. (C, D) Representative Western blots and quantitative data show the amounts of phosphorylated and total AMPK obtained from neutrophils treated with LPS (0-1,000 ng/mL) for 90 min (C) or after exposure to LPS (300 ng/mL) for the indicated time periods (D). Means ± SEM (n = 3), *P< 0.05. (E, F) LPS suppresses activation of AMPK. Representative Western Blots and quantitative data show amounts of phospho and total AMPK, and actin. Neutrophils were treated with AICAR (0 or 250 µmol/L) for 60 min and then cultured with LPS (0 or 300 ng/mL) for an additional 60 min. Cells also were treated with LPS (300 ng/mL) for 60 min followed by inclusion of AICAR (250 µmol/L) in the cultures for an additional 60 min. Means ± SD, n = 3, *P< 0.05, compared with neutrophils treated with LPS alone. ctr, Control; AIC, AICAR.