Toxic Acetaminophen Exposure Induces Distal Lung ER Stress, Proinflammatory Signaling, and Emphysematous Changes in the Adult Murine Lung

Figure 5

Toxic APAP metabolites accumulate in the distal lung. Representative immunofluorescence staining of the distal lung from (a) unexposed and (b and c) APAP-exposed (24 hours; 280 mg/kg, IP) adult male ICR mice. CYP2E1 was stained in red (a), APAP was stained in blue (b), and DAPI was stained in yellow (c). An overlay of all three stains (a, b, and c) is provided (d). Internal scale bar: 50 μm. Flow cytometric analysis demonstrates an induction of CYP2E1 expression in APAP-exposed mice when compared to unexposed controls in (d) epithelial cells and (e) resident alveolar macrophages. (f–k) Flow gating strategy. (f) Scatter cells are gated into (g) singlets, after which (h) neutrophils, B-cells, T-cells, and monocytes are excluded and the Dump^– gate is used to select for (i) epithelial (Dump^-, CD326⁺) or (j) alveolar macrophages (Dump^-, CD45⁺CD64⁺). (k) Resident (Dump^-, CD45⁺CD64⁺Siglec-F^HiCD11b^Lo) and recruited (Dump^–, CD45⁺CD64⁺CD11b^HiSiglec-F^Lo) alveolar macrophages are then segregated based on Siglec-F and CD11b.