Figure 6. Determining if a bacterial mimetic (LPS) can cause the release of EVs in the lung – Signalling.
Mice (n = 6 per group) were challenged with the aerosolised vehicle of endotoxin-free saline or LPS (1 mg/ml) in Perspex chambers for 30 minutes. Animals were sacrificed and BALF obtained 6 hours after challenge. The samples were then centrifuged (900 g) to remove the white blood cells and then pre-treated with inhibitors (P2X7 antagonist A 438079 (10−6 M) or caspase-1 inhibitor VX 765 (10−7 M)) and incubated for 1 hour. Samples were then treated with vehicle (PBS) or ATPγS (10−3 M), and incubated for a further 4 hours and subsequent cytokine release was analysed by ELISA. Data shown as mean +/− S.E.M. (A: IL-1β, B: TNFα). * = P = 0.0138 (One way ANOVA followed by a Bonferroni's Multiple Comparison test).