Extended Data Fig. 3: mCherry⁺-niche neutrophils increase ROS production.

a, b, CD11b⁺ (a) and Ly6G⁺ (b) cell frequencies among live cells in distal lung and mCherry⁺ niche by FACS (n = 9 mice per group). c, Enriched processes by MetaCore analysis and GSEA based on proteomic data by comparing mCherry⁺-niche (n = 3) and distal lung (n = 3) neutrophils; dominant mCherry⁺-niche proteins were obtained by using WebGestalt (http://www.webgestalt.org/option.php). d, PCA of proteins found in unlabelled or mCherry⁺-niche neutrophils (n = 3, each with 10 mice, small circles; large circles represent the average of the triplicates). e, f, Representative FACS plot (e) and scatter plot (f) of intrinsic ROS in Ly6G⁺ cells (n = 6 mice). g, GFP signal quantification of 3D co-culture with GFP⁺ MMTV–PyMT cancer cells and MACS-sorted Ly6G⁺ cells from either naive or metastatic lungs with or without the ROS inhibitor TEMPO (n = 3, each with 3 technical replicates). Data are normalized to cancer cell growth (statistical analysis on biological replicates). h, Representative cancer cell growth on the scaffold (from 14 independent experiments): integrated density of the GFP signal was measured on the scaffold using ImageJ and the corresponding fluorescent image of GFP⁺ cancer cell growth (scale bar, 400 μm). Statistical analysis by paired two-tailed t-test (a, b, f), hypergeometric test with Benjamini–Hochberg correction (c, Metacore), weighted Kolmogorov–Smirnov-like statistic with Benjamini–Hochberg correction (c, GSEA) and two-way ANOVA (g). Data are presented as mean ± s.d. (f) and mean ± s.e.m. (g).

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