Research Paper Volume 14, Issue 19 pp 7692—7717

Natural variation in macrophage polarization and function impact pneumocyte senescence and susceptibility to fibrosis

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Figure 7. NOX2-mediated macrophage superoxide production and senescence inducing capacity varies by strain. (A) Superoxide production was measured by luminol-amplified chemiluminescence in M2 macrophages (IL-13 treated) stimulated with PMA (100 ng/ml) in the presence or absence of a NOX2 inhibitor, GSK2795039 (100 nM). Each symbol: mean, error bars: +SD, *p<0.05 for comparison between macrophages with vehicle and IL-13, §p<0.05 for comparison between macrophages with vehicle +/- GSK2795039, ǂp<0.05 for comparison between macrophages with L-13 +/- GSK2795039 by multiple T Test. (B) Area under the curve analysis for Superoxide production after IL-13 and PMA treatment. (C) Enriched primary AECII from the three strains of mice were seeded on transwell inserts (pore size: 0.4 μm) and cocultured for 3 days with syngeneic M2 macrophages polarized with IL13. AECII were fixed after 3 days, and senescence associated β-gal activity was assessed followed by confirmatory immunocytochemical localization of pro-surfactant C (pro-SPC). The percent of senescent AECII was scored. Columns: mean, error bars: +SD, *p<0.05 for comparison to macrophages with vehicle. §p<0.05 for comparison to C57L macrophages exposed to IL13 by ANOVA with Tukey’s correction.