Figure 6
Peptidoglycan- or lipoteichoic acid-induced NF-κB activation is inhibited by TLR2 blocking antibody in TNF-α-primed and nonprimed HSCs. Activated human HSCs were infected with Ad5NF-κBLuc (MOI 500) for 12 h in DMEM containing 0.5% FCS. HSCs were then incubated with serum-free DMEM or TNF-α (10 ng/ml) for 24 h. After washing three times, HSCs were stimulated with: (a) serum-free media, PGN from S. aureus (10 μg/ml) via preincubation with media, anti-TLR2 bloking Ab (20 μg/ml), or IgG1 Ab for 30 min; or (b) serum-free media, LTA from S. pyogenes (10 μg/ml) via preincubation with media, anti-TLR2 bloking Ab (20 μg/ml), or IgG1 Ab 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 to control HSCs without stimulation; †, §: P<0.01, compared to HSCs with stimulation after preincubation with IgG1 Ab;**: P<0.01, unprimed HSCs vs TNF-α-primed HSCs.
