Functional analysis of host- and donor-derived AMs. (A) Microscopy staining around the alveolus for Siglec F⁺ host-derived (GFP⁻) and donor-derived (GFP⁺) AMs was performed on lead shielded BM reconstituted mice 8-weeks post-CLL delivery. (B) Ratio analysis of host- over donor-derived AMs uptake for apoptotic cell, carboxylated beads, and bioparticles, 2 hours postintranasal delivery. (C) Flow cytometry gating strategy for 4B-4C (left) and ratio analysis of host- over donor-derived AMs for TNF-α and IL-1β production from AM with ingested Escherichia coli (right). Each dot represents one mouse. Blue dots represents analysis from set 1 and red dots represent analysis from set 2 (set 1 and set 2 as described in Figure 1A). (D) Two hours post-IN delivery of bioparticles, AMs were lavaged and quenched to exclude surface bound bioparticles. Black and gray lines are donor- and host-derived AM, dashed is quenched; solid line is unquenched. Control quenching was assessed with ex vivo AM given bioparticles on ice. Blue line is quenched; red line is unquenched. DAPI, 4,6 diamidino-2-phenylindole; GFP, green fluorescent protein.