Extended Data Fig. 2: Alox15−/− AM seeding and maturation.
From: Neonatal imprinting of alveolar macrophages via neutrophil-derived 12-HETE
(a) Representative FACS plot for the determination of AM progenitors (pre-AM and fetal liver monocytes) and AM in the lungs of PND0, PND1 and PND3 WT and Alox15−/− mice. (b) Weight of lungs from PND1 and PND3 WT (n = 7/group) and Alox15−/− mice (n = 6/group). (c-f) Lungs from PND1 or PND3 WT and Alox15−/− mice were harvested and the levels of TGF-β1 (c) (PND1, WT: n = 6, Alox15−/−: n = 7; PND3, WT: n = 8, Alox15−/−: n = 7), GM-CSF (d) (PND1, WT: n = 6, Alox15−/−: n = 7; PND3, WT: n = 8, Alox15−/−: n = 8), type 1 and type 2 cytokines (e- PND1: WT: n = 6, Alox15−/−: n = 7, f- PND3: n = 8/group) were determined by ELISA. (g-h) BAL-AM were isolated from PND3 WT and Alox15−/− mice for gene expression analysis (n = 3 biological replicates/group). (i) BAL-AM were isolated from adult naïve animals and cell death was assessed by flow cytometry using AnnexinV/7AAD staining (n = 7/group). (j-l) Batch effect examinations. (j) PCA and (k) heatmap plots that suggest WT4 is an outlier. (l) A heatmap that indicates the similarity between different samples. Data are presented as mean ± s.e.m and are pooled from two (b-f, i) or three (g-h) independent experiments.
