Figure 5.

cis-NF-κB^EGFP transcriptional induction in F4/80⁺ hepatic macrophages at 4, 24 h after in vitro LPS challenge prior to chronic ethanol feeding. Livers of cis-NF-κB^EGFP mice were harvested 4 weeks after Lieber-DeCarli diet treatment and Kupffer cells were purified and cultured as described in Section 2. Cells were then stimulated with 10 μg/mL LPS over a period of 24 hours. Red fluorescence (F4/80) labels Kupffer cells and green fluorescence of EGFP identifies cells expressing NF-κB transcriptional activity at 4, 24 h after LPS stimulation. (a) and (b) show representative unstimulated controls whereas in (c) and (d) representative overlay images at 4 h after LPS stimulation are given (magnification, ×200). Colabeled (yellow) cells and F4/80 positive (red) cells were counted. The percentage of F4/80⁺EGFP⁺ cells in total F4/80⁺ cells is depicted in (e) at 4, and 24 h after LPS stimulation. Data shown are representative of five to eight separate experiments and are presented as mean ± SEM. *P < 0.05 versus Ctrl.