Extended Data Fig. 9: 12-HETE-treated pups have restored proliferation.

(a) Alox15 expression was measured in BAL AM (PND3 and adult mice) or peritoneal macrophages using qPCR (n = 5/3/3 biological replicates/group). (b) Pulmonary levels of 15(S)-HETE in PND1, PND3 and adult WT mice (n = 10/8/8/group). (c) Representative FACS plots and quantification of Ki67⁺ AM after 12-HETE treatment (Veh.: n = 7, 12-HETE: n = 5). (d) AM populations after 12-HETE treatment (Veh.: n = 7, 12-HETE: n = 6). (e) Representative FACS plots and quantification of Ki67⁺ AM after 15-HETE treatment (Veh.: n = 3, 15-HETE: n = 4). (f) Representative micrographs of BrdU staining in vitro after AM culture with GM-CSF. Bar=50 µm. (g-i) PND1 WT pups were delivered intranasally anti-FcεR1 antibodies or isotype controls for two consecutive days. (g) Basophil depletion, (h) AM populations and (i) Ki67⁺ AM were determined at PND3 (Iso.: n = 4, α-FcεRI: n = 6). (j) Representative histograms of ALOX15 expression in CD45⁺ or CD45⁻ lung cells at various ages of WT and Alox15^−/− mice. (k) PND1 ALOX15⁺ CD45⁺ cells were further gated using CD11b, Ly6C and Ly6G. (l) Lung and blood neutrophils were isolated from PND1 or adults (6–8 weeks) WT or Alox15^−/− mice. Expression of ALOX15 and Ly6G were assessed by immunofluorescence. Blue: DAPI. Bar = 20 µm. (m) Representative micrographs of ALOX15 and Ly6G expression in frozen lung sections of PND1 WT pups. Data are presented as mean ± s.e.m and are from one (g-i) or pooled from two (c-e) or three (a-b) independent experiments or representative from two (f, j, k, l, m) independent experiments. Data were analyzed using two-tailed unpaired t-test (c, g) or one-way ANOVA followed by Tukey’s multiple comparisons test (b).

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