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. Author manuscript; available in PMC: 2011 Jun 20.
Published in final edited form as: Thorax. 2010 Apr;65(4):346–353. doi: 10.1136/thx.2008.101139

Figure 1. SPR activation inhibits IFN-γ-mediated NO production, iNOS mRNA and protein in alveolar macrophages.

Figure 1

Figure 1

Panel A. Heat stress inhibits IFN-γ-mediated NO release in the extracellular medium in a time-dependent manner. MH-S cells were incubated at 43°C for increasing times, then allowed to recover at 37°C for 1 hour prior to IFN-γ stimulation (10 ng/ml) for 24 hours. Extracellular medium was assayed for nitrite using the Griess reagent. Panel B Heat stress does not affect the viability of MH-S cells. MH-S cells were incubated in 10% Alamar Blue reagent diluted in cell medium and incubated at 37°C for 2 hours. Color change was determined spectrophotometrically. For both panels, results are the mean ± SD of 4 separate experiments with each condition carried out in triplicate. Panel C. Heat stress inhibits IFN-γmediated iNOS mRNA expression. MH-S cells were either untreated or were incubated at 43°C and allowed to recover for 1 hour prior to IFN-γ stimulation (10 ng/ml for 2h) and analysis for iNOS mRNA by real-time RT-PCR normalized with GAPDH mRNA levels. Results are the mean ± SD of three experiments done in triplicate; *p < 0.05 from control cells; **p < 0.05 from cells treated with INF-γ alone. Panel D. Heat stress inhibits IFN-γ-mediated iNOS protein expression in MH-S cells. MH-S cells were either untreated or were incubated at 43°C and allowed to recover for 1 hour prior to IFN-γ stimulation (10 ng/ml for 7h) and analysis for iNOS protein by Western blot; one representative experiment is shown, three additional experiments gave comparable results.