Extended Data Fig. 6: Homeostatic Alox15^−/− AM are similar to WT AM.

(a) Heatmap and quantification of genes previously associated with alveolar macrophages or monocytes lineages (WT: n = 3, Alox15^−/−: n = 4). (b) Representative FACS plots and quantification of surface markers expression determined by flow cytometry. (c) Heatmap and quantification of genes previously associated with alveolar macrophages phenotype (WT: n = 3, Alox15^−/−: n = 4). (d) Representative FACS plots of AM population in 6–8 weeks and 52 weeks old WT and Alox15^−/− mice. (e) AM numbers in the lungs of 6–8 (WT: n = 3, Alox15^−/−: n = 5) or 52-weeks old (n = 5/group) WT and Alox15^−/− mice. (f) BAL total protein levels in 6–8 (n = 3/group) or 52-weeks old (n = 5/group) WT and Alox15^−/− mice. (g) Representative micrographs of lung sections from naïve 6–8 weeks and 52 weeks old WT and Alox15^−/− mice. Bar = 100 µm. (h-i) Adult WT (n = 5) and Alox15^−/− (n = 6) mice were used to measure resistance (h) and elastance (i). (j) Phagocytic capacity of naïve BAL AM from adult WT and Alox15^−/− mice (WT: n = 7, Alox15^−/−: n = 6 biological replicates). (k) Representative micrographs of BAL AM, cytospun, and stained with H&E (top) or Oil red O (bottom). Bar = 20 µm. (l-n) BAL was collected from adult WT, Alox15^−/− and Csf2rb^−/− mice, and BAL turbidity (l) (n = 10/14/11/group), total protein content (p) (n = 13/15/15/group) and SP-A levels (q) (n = 5/6/5/group) were measured. Data are presented as mean ± s.e.m and are from one (a, c, d-f) or pooled from two (h-j) or three (l-m) independent experiments or representative of two (b, k) independent experiments or 5 biological replicates (g). Data were analyzed using two-way ANOVA followed by Tukey’s multiple comparison test (f), two-tailed unpaired t-test (e), or one-way ANOVA followed by Tukey’s multiple comparisons test (l-n).

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