Altered polarization of PAR-2 signaling during airway epithelial remodeling

Ryan M. Carey; Jenna R. Freund; Benjamin M. Hariri; Nithin D. Adappa; James N. Palmer; Robert J. Lee

If primary cells were incubated for a further 4 days with the apical side submerged (Day 21 + 4 submerged) to promote squamous de-differentiation, the response to apical 2FLI was again observed (Fig 3C and F) . Basolateral application of antagonist FSLLRY-NH2 blocked the response to basolateral but not apical 2FLI (Fig 3C, right) , ruling out 2FLI diffusion across the epithelial barrier to the basolateral side. Similar results were seen at ALIs after 4 subsequent days of basolateral IL-13 (Fig 3D) . If primary ALIs were carried out to day 25 with no IL-13 and no apical submersion, only basolateral 2FLI responses were observed (Fig 3F) . RPMI2650 squamous cells cultured at ALI also had 2FLI responses with application to either the basolateral or apical side; basolateral addition of antagonist blocked basolateral but not apical responses (Fig 3E and F) . No significant differences in TEERs were observed (Fig 3G) .