Fig. 2: Alox15−/− AMs are intrinsically impaired in proliferation.
From: Neonatal imprinting of alveolar macrophages via neutrophil-derived 12-HETE
a, Representative FACS plots (left) and quantification (right) of BrdU+ AMs in adult WT and Alox15−/− lungs after a 7-day BrdU pulse (n = 5 mice per group). b, Mouse Ki67 (mKi67) expression from the RNA-seq dataset (n = 3 (WT) or 4 (Alox15−/−)). c, Representative FACS plots (left) and quantification (right) of Ki-67+ AMs in adult WT and Alox15−/− lungs (n = 11 per group). d, Scheme of the adoptive transfer protocol. e, Ki-67+ AMs before and after transfer (WT CD45.1 (left to right), n = 4, 5 or 5; Alox15−/− CD45.2, n = 5 per group). f, Growth of AMs after culture with GM-CSF for 3 days (n = 4 biological replicates per group). g,h, Representative micrographs (left) and quantification (right) of BrdU+ (g) (n = 3 (unstimulated (Uns.) or 5 (GM-CSF) fields of view) and Ki-67+ (h) (n = 5 (Uns.), 12 (WT GM-CSF) or 10 (Alox15−/− GM-CSF) fields of view) AMs after 3 days of culture with GM-CSF. Scale bar, 50 µm. i, Representative FACS plots (left) and quantification (right) of Ki-67+ AMs in PND3 lungs (n = 12 mice per group). j, Scheme of ATAC-seq of AMs from adult (BAL) or PND3 (sorted) WT and Alox15−/− mice (n = 3 mice per group). k,l, Volcano plots for differential accessibility results in adult (k; knockout (KO) versus WT) and pups (l). Yellow (k) or blue (l) highlighted peaks have a P-adjusted value of <0.05 and absolute log2(fold change (FC)) of ≥1. m, The log2(FC) difference in accessibility in adults and pups for all significant peaks within pups. Pink highlighted points are DA in both adults and pups. Green points are significant only in pups. n, Top ten enriched pathways in adults and pups from a GSEA of genes matched to the closest DA peaks. Data are presented as the mean ± s.e.m. and are from one (e,k–n) or pooled from three (c,i) or four (f) independent experiments or representative of two (a) or three (g,h) independent experiments. Data were analysed using unpaired two-tailed t-test (a–b,i) or two-way ANOVA followed by Sidak’s (e) or Tukey’s (f–h) multiple comparisons test. The models in d and j were created using BioRender (https://biorender.com).
