Extended Data Fig. 6: A-Eos are induced by NF-κB signalling.

a, Regulon activity across clusters. b, Representative regulons projected on UMAP plot. Cells are coloured by binary regulon activity. c, Pathway activity across clusters according to PROGENy analysis. d, Gene expression relative to Hprt measured by qRT–PCR of eosinophils sorted from the blood (n = 6), spleen (n = 6) and colon (n = 4) of Il5-tg mice. Data represents mean ± SEM. One-way ANOVA. e, Representative images of pNF-κB p65 immunofluorescence staining in colonic eosinophils (n = 3, B6J). Arrows mark A-Eos (Siglec-F⁺ CD80⁺, red) and B-Eos (Siglec-F⁺ CD80⁻, green). Nuclei are stained with DAPI. Scale bar, 20 µm. f, A-Eos frequencies upon conditioning of BM-Eos with colon CM and/or NF-κB inhibitor (n = 5, B6J). Data represents mean ± SD. One-way ANOVA. g, Representative FACS plots of colonic A-Eos (PD-L1⁺CD80⁺) and PD-L1⁻CD80⁻ (B-Eos), relative to Fig 3d. Numbers indicate % of eosinophils. h, Left: Representative FACS plots of colonic A-Eos (PD-L1⁺CD80⁺) and PD-L1⁻CD80⁻ (B-Eos), relative to Fig 3e. Numbers indicate % of eosinophils. Right: MFI of Siglec-F and % CD63 in colonic A-Eos and B-Eos shown in Fig. 3e. Medians are shown. Two-tailed unpaired Student’s t-test. i, Colonic A-Eos and B-Eos frequencies at steady state in B6J (n = 5) Tlr2^−/− (n = 3) and Tlr4^−/− (n = 7) mice. Medians are shown. One-way ANOVA.