Fig 2.

HMGB1 mRNA and protein expression in ASMCs assessed by (A) quantitative PCR and flow cytometry (B) at baseline with representative histograms to the right and (C) following stimulation with proinflammatory cytokines (TNF-α, IFN-γ, and IL-1β, 10 ng/mL) or the dsRNA mimic poly(I:C) (12.5 μg/mL). *P < .05 versus unstimulated ASMCs. ROS production in response to (D) HMGB1 (10-1000 ng/mL) in ASMCs, *P ≤ .05 healthy control (●) vs patient with asthma (■), unpaired t test, (n = 5-9 ASM donors), and (E) HMGB1 (1000 ng/mL) ± LPS-RS or sRAGE (10 μg/mL) in ASMCs from healthy controls. P values from unpaired t tests. F-H, Contraction of collagen gels impregnated with ASMCs from subjects with asthma in the presence of bradykinin following incubation with vehicle control or 1000 ng/mL HMGB1 ± LPS-RS or sRAGE (10 μg/mL) expressed as (Fig 2, F) time course, *P < .05 vs vehicle control, (Fig 2, G) example collagen gels at 60 minutes, and (Fig 2, H) AUC at 60 minutes. *P < .05 vs vehicle control, paired t tests. Means ± SEM are shown. GMFI, Geometric mean fluorescence intensity; PE, phycoerythrin.