Figure 4

Peptidoglycan or lipoteichoic acid induces NF-κB transcriptional activation in activated human HSCs. NF-κB transcriptional activation was assessed with NF-κB-driven luciferase assay. Activated human HSCs were infected with Ad5NF-κBLuc (MOI 500) for 12 h in DMEM containing 0.5% FCS. At 20 h post-infection, HSCs were stimulated with PGN (S. aureus) (10 μg/ml), PGN (B. subtilis) (10 μg/ml), LTA (S. pyogenes) (10 μg/ml), TNF-α (10 ng/ml), or LPS (1 μg/ml) for 8 h in serum-free conditions with or without pretreatment of Polymyxin B (10 IU/ml) for 30 min. Cells were lysed, and NF-κB-mediated luciferase activity was quantified. Data represent the mean±s.d. of three independent experiments performed in duplicate, and are expressed as fold-increase over unstimulated cells. All measurements of luciferase activity were normalized to the protein concentration. ^*, †: P<0.01, when compared with unstimulated HSCs.