SERCA2 Regulates Non-CF and CF Airway Epithelial Cell Response to Ozone
Figure 3
Effect of ozone exposure on the release of proinflammatory cytokines in differentiated cultures of non-CF (▵, 0 ppb and ▴, 200 ppb) and CF (○, 0 ppb and •, 200 ppb) primary airway epithelial cells. Cells were cultured on collagen-coated snapwells and allowed to grow and differentiate for 30 days. Before ozone exposure 100 µl media was added to the apical surface. At the end of exposure (18 h) an additional 200 µl media was added apically. After 4 h the media from apical (A) and basolateral (B) surface was collected and 100 µl of 6 mM DTT in PBS was added on the apical surface. The cells were incubated for 15 min in DTT and the supernatant fluid was carefully collected using a 1.0 ml pipette tip. The media and DTT solubilized mucin were analyzed for cytokines (IL-8 in A & B, G-CSF in C & D and GM-CSF in E & F) as described in the Methods. For apical cytokine content media and DTT solubilized mucin samples were pooled. Each symbol represents the mean of each donor. Three separate experiments were performed each time using one non-CF (donor 1-3) and one CF (4-6) donor. * Indicates significant difference from 0 ppb exposed non-CF, p<0.05 and # indicates significant difference from 200 ppb non-CF, p<0.05 and $ indicates significant difference from 0 ppb CF, p<0.05.