Fig. 4: γδ17 rendered neutrophils to present a more pro-tumoral phenotype in an aging state.

Proportions of CXCR2+ cells in neutrophils in bone marrow from AC/YC groups (a) and AM/YM groups (b), measured by flow cytometry, gated on CD45+CD11B+LY6G+ cells. Each group contains six mice. Data expressed as % of CXCR2+ neutrophils (mean ± SD). P = 4.8E−03 (a); P = 2.1E−05 (b). Significance was determined using unpaired two-tailed Student’s t test. **P < 0.01, ****P < 0.0001. c Based on the scRNA-seq data from lung, heatmap showing representative KEGG pathways of neutrophils across AC, YC, AM and YM groups, calculated by GSVA packages. Color key from white to red indicates expression levels from low to high. d Proportions of lung c-Kit+ neutrophils were measured by flow cytometry, gated on CD45+CD11B+LY6G+ cells. Each group contains six mice. Data expressed as % of c-Kit+ neutrophils (mean ± SD). YC-AC (P = 0.01), YM-AM (P = 1.0E−04). Significance was determined using one-way ANOVA. *P < 0.05, ***P < 0.001, ns P > 0.05. e Dot plot showing representative GO terms and KEGG pathways enriched in upregulated DEGs of neutrophils in the AC/YC and AM/YM comparison. Significance was calculated based on the accumulative hypergeometric distribution by Metascape webtool. f MFI of CXCR4 in neutrophils of lung from AM/YM/AC/YC groups, which were measured by flow cytometry, gated on CD45+CD11B+LY6G+ cells. FMO fluorescence minus one control. Each group contains six mice. Data expressed as MFI of CXCR4 (mean ± SD). YC-YM (P = 8.4E−03), AC-AM (P = 1.1E−06), YM-AM (P = 3.5E−04). Significance was determined using one-way ANOVA. **P < 0.01, ***P < 0.001, ****P < 0.0001, ns P > 0.05. Representative images and quantitative results of lung metastases in aged (g) and young (h) mice with anti-γδTCR or isotype (control) treated. Each group contains six mice. Data expressed as number of metastases (mean + SD). P = 0.01 (g); P = 0.7 (h). Significance was determined using unpaired two-tailed Student’s t test. *P < 0.05, ns P > 0.05. Proportions and quantity of neutrophils (i) and c-Kit+ neutrophils (j) were measured by flow cytometry, gated on CD45+ cells or CD45+CD11B+LY6G+ cells. Each group contains six mice. Data expressed as % or number of neutrophils, c-Kit+ neutrophils (mean ± SD). Neutrophils-ratio: AM (P = 2.3E−05), YM (P = 6.5E−03); Neutrophils-number: AM (P = 1.4E−04), YM (P = 0.03). c-kit+ neutrophils-ratio: AM (P = 6.74E−03), YM (P = 0.93); c-kit+ neutrophils-number: AM (P = 1.8E−03), YM (P = 0.08). Significance was determined using unpaired two-tailed Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns P > 0.05. Source data are provided as a Source Data file.