Figure 6

Cytokine-induced p38 MAP kinase activation does not augment subsequent TLR2 stimulation. A) Phosphorylated and total p38 cellular protein levels were assessed using immunoblot analysis of extracts from human airway epithelia that were treated without or with media containing TNF-α (100 ng/ml) and IFN-γ (100 ng/ml) for 30 min. B) Phosphorylated and total p38, and heat shock protein-90 (HSP90) cellular protein levels were assessed using immunoblot analysis of extracts from human airway epithelia that were first treated without or with media containing TNF-α (100 ng/ml) and IFN-γ (100 ng/ml) for 24 hours, followed by incubation without or with Pam3CSK4 (25 mg/ml) for 30 min. C) Phosphorylated p38 and HSP90 protein levels in the experiment outlined in B were quantified using band densitometry of immunoblot analyses results with inclusion of samples from three individuals. Values were calculated as phosphorylated p38/HSP90, were normalized to the untreated control value for each individual, and are expressed as mean fold change in phosphorylated p38 ± S.D. (n = 3 in each group).