Fig. 4

Flow cytometric quantification of LLP2A binding to different lung leukocytes. Representative histograms from flow cytometry of enzymatically dissociated murine lung single cells (A) demonstrate negligible binding of LLP2A-Biotin to CD45-negative cells (top panel) while different levels of binding to LLP2A-Biotin are present in CD45-positive leukocytes (bottom panel). The specificity of LLP2A-Biotin binding is confirmed by near-complete blocking of its binding in cells co-incubated with excess non-fluorescent LLP2A. Cell-type specific analysis of the flow cytometry data demonstrated that LLP2A-Biotin binding nearly doubles in alveolar macrophage (aMφ) of bleomycin-injured lungs compared to control lungs, while is not significantly different compared to control lungs in the remaining leukocyte subsets (B). There is a significant increase in the abundance of interstitial and monocyte-derived macrophage (iMφ & MDMφ) as well as dendritic cells (DC) at 1-week post-bleomycin injury versus controls (C). N = 5 (control group) and 6 (bleomycin group)