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. Author manuscript; available in PMC: 2018 Oct 15.
Published in final edited form as: J Immunol. 2017 Sep 8;199(8):2873–2884. doi: 10.4049/jimmunol.1700539

Figure 8. Neutrophil-derived cPLA2α plays a key role in migration across a human primary airway mucosal model.

Figure 8

A) μOCT imaging displays trans-epithelial migration of neutrophils across a human airway basal stem cell-derived epithelium, grown in air-liquid interface. B) Neutrophils were pretreated for 60 min. with cPLA2α inhibitor (12μM) or vehicle control (DMSO 1:500) then allowed to migrate across a PAO1 infected mature epithelium derived from human airway basal stem cells grown in air-liquid interface. Migration was visualized by micro-OCT imaging. Representative images across the 2h migration period were selected. C) Neutrophil migration was quantified by neutrophil density per area (mm2) and plotted over time. D) Migration of neutrophils pretreated for 60 min. with cPLA2α inhibitor (12μM) or control (DMSO 1:500) across a mature human airway basal stem cell-derived epithelium in response to PAO1 infection or buffer (HBSS) was quantified by total myeloperoxidase activity. E) After the 2h migration, supernatant was collected from the apical compartment and LTB4 was quantified by ELISA. F) In the human airway basal stem cell-derived ALI model, LTB4 production was corrected for by neutrophil migrated and compared to display relative LTB4 production per neutrophil. Data are shown as mean corrected value +/− standard deviation. Experiments were performed on at least three occasions with n≥3 technical replicates. P values were calculated by paired Student’s t-test. P values ≤0.05 was consider significant.