SiglecF ^hi TANs secrete TGFβ. (A) GO analysis of differentially expressed genes between RIL175 cells and those co-cultured with SiglecF^hi TANs. BP, biological process. (B and C) NicheNet analysis of scRNA-seq data from NRAS/AKT HCC mice treated with αLy6G or isotype control mAbs. Prioritized ligands and their target genes driving transcriptomic changes in tumor cells after αLy6G treatment (B). Log fold-change (LFC) in senders of ligands to tumor cells after αLy6G treatment (C). (D) Relative expression of indicated genes among neutrophil subsets from NRAS/AKT HCC mice. (E) Representative FACs plots (left) and percentage of LAP (TGFβ1)⁺ cells among TAN subsets. (F) Spontaneous TGFβ production by SiglecF^– and SiglecF^hi TANs. Each symbol represents one mouse (E) or TANs isolated from two to three mice (F). Data are mean ± SEM and represent two independent experiments (E and F). *P < 0.05, ****P < 0.0001 by unpaired Student’s t test (E and F).