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. 2016 Sep 30;11(9):e0163751. doi: 10.1371/journal.pone.0163751

Fig 2. IL-33 mediates eosinophil survival via autocrine GM-CSF production.

Fig 2

(A) BMDE were cultured in medium containing 10 ng/ml IL-5 and indicated concentrations of IL-33 and LPS for 24 hours. Supernatants were analyzed by GM-CSF-specific ELISA. Bar graph shows the mean + SD from one of two independent experiments (n = 4;). (B) BMDE were stimulated with the indicated cytokines in the absence (black bars) or presence of a isotype control antibody (dark grey bars) or GM-CSF receptor blocking antibody (light grey bars). After 72 hours cells were analyzed by flow cytometry to detect Annexin V+ cells. Bars show the mean + SEM of apoptotic cells from three independent experiments (n = 6; ** p<0.01). (C) Sort-purified BMDE (100% purity, see S1 Fig) were left untreated (black bar) or stimulated with IL-33 in the presence of isotype control antibody (dark grey bar) or anti-GM-CSF-R antibody (light grey bar). After 72 hours cells were analyzed by flow cytometry to detect Annexin V+ cells. Bars show the mean + SEM of apoptotic cells from two experiments (n = 4; * p<0.05; ** p<0.01). (D) BMDE were treated as described above and harvested after 24 hours to analyze Bcl-xL transcripts by quantitative RT-PCR. Bar graphs show mean + SEM Bcl-xL mRNA expression levels normalized to HPRT1 mRNA levels with data pooled from two independent experiments (n = 6; * p<0.05).