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. 2016 Jul;55(1):58–71. doi: 10.1165/rcmb.2015-0183OC

Figure 1.

Figure 1.

Signal transducer and activator of transcription (STAT) 6 is essential for Cu,Zn–superoxide dismutase (SOD)–mediated M2 polarization. (A) Wild-type (WT) and STAT6−/− bone marrow–derived macrophages (BMDMs) were cultured in the presence or absence of polyethylene glycol (PEG)–conjugated SOD (PEG-SOD) for 3 hours and then treated with chrysotile asbestos. Total RNA from macrophages was isolated, and tnf-α, chitinase-like 3 (ym1), and resistin-like molecule alpha (fizz1) gene expression were measured by quantitative RT-PCR. Results show arbitrary units normalized to β-actin mRNA (n = 3). *P < 0.05 WT versus STAT6−/−, **P < 0.05 WT versus WT + PEG-SOD, #P < 0.05 STAT6−/− + PEG-SOD versus STAT6−/−. (B) Macrophages were transfected with either scrambled or STAT6 small interfering RNA (siRNA) for 72 hours. Total mRNA was isolated and chemokine ligand (ccl-18) gene expression was measured. Results show arbitrary units normalized to hypoxanthine-guanine phosphoribosyltransferase mRNA (n = 3); *P < 0.05 STAT6 siRNA versus scrambled siRNA. (C) WT and STAT6−/− macrophages were stained with a fluorescent compound (R-PE)–labeled anti-CD23 at a 1:10 dilution. Levels of CD23 were determined by flow cytometry. Representative of three histograms of CD23-stained macrophages is shown. WT and STAT6−/− mice were exposed to chrysotile (100 μg) intratracheally. (D) Ym1 (n = 5) and (E) TNF-α (n = 3) in the bronchoalveolar lavage (BAL) fluid were measured 21 days after chrysotile exposure; *P < 0.05. (F) Total RNA from alveolar macrophages was isolated, and tnf-α, ym1, and fizz1 gene expression were measured as in A. Results show arbitrary units normalized to β-actin mRNA (n = 3); *P < 0.05 versus WT.