IL-33 expression induced by <i>S.aureus</i>-derived PGN and LTA in macrophages.

<p>(<b>A</b>&<b>B</b>) Quantification of IL-33 expression in macrophages RAW264.7 treated with different doses of PGN (<b>A</b>) or treated with 10 µg/ml PGN for different times (<b>B</b>). (<b>C</b>&<b>D</b>) Quantification of IL-33 expression in macrophages RAW264.7 treated with different doses of LTA (<b>C</b>) or treated with 10 µg/ml LTA for different times (<b>D</b>). (<b>E</b>) Western blot of IL-33 induced by commercial PGN (cPGN) and PGN purified from <i>S.aureus</i> CMCC(B)26003 (pPGN) in total cell lysate of primary peritoneal macrophages. (<b>F</b>) Western blot of IL-33 induced by PGN in total cell lysate of primary peritoneal macrophages at different time points. (<b>G</b>) Western blot of IL-33 from the cytoplasm or nucleus of macrophages RAW264.7 treated with 10 µg/ml PGN. Histone was used as an endogenous control for nuclear proteins and tubulin was used as an endogenous control for cytoplasmic proteins. (<b>H</b>) Immunofluorescent staining of IL-33 in macrophages RAW264.7 treated with PGN for 24 hours. Scale bar represents 50 µm. (<b>I</b>) IL-33 in cell culture media. Cell culture media of macrophages RAW264.7 treated with 10 µg/ml PGN was incubated with protein G beads coupled with IL-33 antibody for overnight. Next day, IL-33 captured by IL-33 antibody was eluted from beads for western blot. *<i>P<0.05</i> and <i>***P<0.001</i>. <i>P</i> values were determined by one-way ANOVA. Data are the means ± SEM of <i>n</i> = 3 and representative of three independent experiments.