Mediators of Inflammation / 2014 / Article / Fig 7

Research Article

Effect of Tumor Necrosis Factor Family Member LIGHT (TNFSF14) on the Activation of Basophils and Eosinophils Interacting with Bronchial Epithelial Cells

Figure 7

Effects of signaling molecule inhibitors on the cell surface expression of ICAM-1 and induction of cytokines/chemokines from coculture of BEAS-2B cells and basophils/eosinophils in the presence of LIGHT (100 ng/mL). Cocultures of (a, b, c, d) basophils or (e, f) eosinophils (3 × 105 cells) and confluent BEAS-2B cells (1 × 105 cells) were pretreated with BAY11-7082 (1 μM), SP600125 (3 μM), SB203580 (7.5 μM), U0126 (10 μM), or LY294002 (10 μM) for 1 h, followed by incubation with or without LIGHT (100 μg/mL) in the presence of inhibitors for a further 24 h. Cell surface expression of ICAM-1 on 5,000 cells was analyzed by flow cytometry as MFI. Results are expressed as mean plus SEM of three independent experiments with three blood samples in bar charts. Release of cytokines/chemokines and MMP-9 in culture supernatant was measured by Milliplex assay. Results are expressed as mean plus SEM. DMSO (0.1%) was used as the vehicle control. * , ** when compared with vehicle or negative control.

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