Fig. 4: Signals that drive neutrophil maturation along the different paths.
a, In silico screening of cytokine induction of transcriptional profiles25. Heat map shows lymph node-derived neutrophils treated in vivo with cytokines or PBS (arrowhead). Cells were classified into NeuMap hubs using Seurat LabelTransfer; proportions are colour-coded by hub. b, Distribution of transcriptomes from cytokine-treated neutrophils projected onto NeuMAP. c, Heat map of 21 markers in bone marrow-derived neutrophils (left) and UMAP projection (right) defining five states (immature, mature, inflammation/infection, IS-I and IS-II). Data from n = 4 independent experiments. d, Contour plots of neutrophils from c after 24 h treatment with cytokines or conditioned media from LLC (CM LLC) and PDAC (CM PDAC). e, Radar chart summarizing neutrophil distributions from d. f, Scheme, haematoxylin and eosin (H&E) staining and NeuMAP projection (K-mass) of bone marrow neutrophils cultured with TGFβ, IFNβ or GM-CSF. Micrographs show representative nuclear morphologies (immature ringed versus multilobulated). Scale bars, 10 µm. g, Heat map visualization of the proportion of bone marrow neutrophils from TgfbrΔN, IfnarΔNand Csf2rΔN mutants and Cre− controls in the mature, inflammation/infection (infl./infect.) and cancer phenotypic clusters after treatment with the indicated cytokines for 24 h, determined by flow cytometry. Data are mean ± s.e.m. from n = 3 biologically independent mice per group. One-way ANOVA followed by Dunnett’s multiple comparison test. h, Contour plots showing distribution of neutrophils from indicated genotypes in NeuMAP; shifts (arrowheads) are quantified in Extended Data Fig. 8f. i, Functional assays of neutrophils treated with vehicle or cytokines, measuring migration (n = 3–4), bacterial killing (n = 6–7), phagocytosis (n = 4), NET formation (n = 10–13), immunosuppression (n = 4–6) and angiogenesis (n = 7–8). Data are mean ± s.e.m. from n = 2 independent experiments. One-way ANOVA followed by Dunnett’s multiple comparison test. FC, fold change.
