Fig. 6

LIT-927 impedes pericyte uncoupling from the airway microvasculature. Female C57/Bl6 mice (6–8 weeks old) were subjected to intranasal delivery of sterile PBS (10 µl) or house dust mite extract (HDM; 25 µg in 10 µl) 5 days a week for 5 consecutive weeks. A, B At the end of the protocol, the trachea and bronchi were collected, cleaned, and stained as a whole mount to perform a three-dimensional analysis of uncoupled pericytes mobilized, uncoupled pericytes (pericytes that have dissociated from the microvasculature). Tracheobronchial whole mounts were stained for the mesenchymal cell marker α-smooth muscle actin (α-SMA; red) and the endothelial cell marker CD31 (cyan). Scale bar 25 µm (A) and 10 and µm (B). C Uncoupled α-SMA+ cells (example indicated by black arrows) were counted and D the area:perimeter ratio was calculated. n = 5–8 per group from two independent experiments; *p < 0.05. E Lungs were processed into a single cell suspension and submitted to flow cytometric analysis to determine the number and proportion of CXCR4 + pericytes. n = 7 per group, representative of two independent experiments; *p < 0.05, **p < 0.01. HDM: house dust mite; LIT: LIT-927 (CXCL12 neutraligand); PBS: phosphate-buffered saline; VEH: vehicle control