Extended Data Fig. 5: Proliferation of tumour cells adjacent to neutrophils in primary and metastatic tissues.

a, Representative immunofluorescence images of NSG-LM2-GFP primary tumour and matched lung metastasis, stained for pan cytokeratin (pCK, green), MPO (gold), Ki67 (purple) and DAPI (nuclei, blue) (n = 3). b, Plots showing the mean per cent of Ki67-positive cancer cells in the primary tumour and metastatic sites (lung or brain) of mouse models, both overall and when considering only those cells that are adjacent to neutrophils. n = 3 for all; error bars, s.e.m.; ns, not significant by two-sided Student’s t-test. c, Representative immunofluorescence images of BR57 primary tumour and matched liver metastasis, stained for pan cytokeratin (pCK, green), MPO (gold), Ki67 (purple) and DAPI (nuclei, blue) (n = 3). d, Plots showing the mean percentage of Ki67-positive cancer cells, both overall and when considering only those cells that are adjacent to neutrophils, in matched primary and metastatic sites of nine patients with breast cancer. Error bars, s.e.m.; ns, not significant by two-sided Student’s t-test. e, Schematic of the experimental design. One hundred CTCs from CTC–neutrophil clusters or CTC alone are injected in the tail vein of recipient mice to measure disseminated tumour cells proliferation. f, Representative images of disseminated tumour cells stained for pan-cytokeratin (pCK, green), Ki67 (purple) and DAPI (nuclei, blue) (left). The plot shows the mean per cent of Ki67-positive DTCs (right). n = 3; error bars, s.e.m.; ^*P = 0.001 by two-sided Student’s t-test.

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