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Figure 5 | Respiratory Research

Figure 5

From: Inhibition of IFN-γ-dependent antiviral airway epithelial defense by cigarette smoke

Figure 5

Cigarette smoke extract decreases type II interferon-dependent antiviral defense. A: RSV N-gene mRNA levels were determined using realtime RT-PCR analysis of total RNA from hTBE cells that were first treated with media without or with CSE at the indicated concentration for 4 hours, followed by incubation for 20 hours in media containing the same CSE concentration without or with IFN-γ. Cells were then infected with RSV for 24 hours. Values are expressed as mean normalized N-gene mRNA level compared to control HPRT mRNA level (n = 3-4 experiments), and a significant difference (p < 0.05) in untreated or CSE-treated levels between cells uninduced versus induced by IFN-γ is indicated by an asterisk. B: RSV and β-actin protein levels were assessed using immunoblot analysis of extracts from hTBE cells that were first treated with media without or with CSE at the indicated concentration for 4 hours, followed by incubation for 8 hours in media containing the same CSE concentration without or with IFN-γ. Cells were then infected with RSV for 24 hours. C: Protein levels were quantified using band densitometry of immunoblot analyses from separate experiments performed as outlined for B. The band intensity of RSV G/β-actin was set at 1 in each experiment for cells not exposed to CSE or IFN-γ and other conditions were compared relative to this value. Values are expressed as mean ± S.D. (n = 4 experiments), and a significant difference (p < 0.05) in untreated or CSE-treated levels between cells uninduced versus induced by IFN-γ is indicated by an asterisk.

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