Extended Data Fig. 4: Normalizing the NK cell pool prevents hepatic metastases.
From: Hepatic stellate cells suppress NK cell-sustained breast cancer dormancy
a, Flow cytometry quantification of NK cell frequency in mice treated with IgG, anti-GM1, PBS or IL-15. Left, MDA-MB-231 model (n = 8 IgG, n = 10 anti-GM1, n = 8 PBS, n = 10 IL-15; data combine two independent experiments). Right, 4T1 model (n = 4 IgG, n = 5 anti-GM1, n = 5 PBS, n = 5 IL-15). b, Bioluminescence imaging 10 weeks after MDA-MB-231 tumour resection. c, d, Quantification of metastatic foci (c) and metastatic area (d) in livers of mice treated with IgG, anti-GM1, PBS or IL-15, normalized to the liver lobe area analysed. Left, MDA-MB-231 model (n = 8 IgG, n = 10 anti-GM1, n = 8 PBS, n = 10 IL-15; data combine two independent experiments). Right, 4T1 model (n = 5 IgG, n = 10 anti-GM1, n = 6 PBS, n = 10 IL-15). e, Experimental design for examining the effects of NK cell depletion on the 4T07 model. f, Sustained NK cell depletion reactivates dormant 4T07 DTCs in the liver. Arrowheads indicate single Ki67− quiescent DTCs. Coloured line delineates a metastasis. Scale bars, 30 μm. g, Quantification of liver metastatic foci after NK cell depletion in the 4T07 model, normalized to the liver lobe area analysed (n = 10 IgG, n = 10 anti-GM1). h, Quantification of scattered quiescent DTCs in livers of mice treated with IgG, anti-GM1, PBS or IL-15, normalized to the liver lobe area analysed (n = 8 IgG, n = 10 anti-GM1, n = 8 PBS, n = 10 IL-15; data combine two independent experiments). i, Expression of IL-15Rα in MDA-MB-231, 4T07 and 4T1 cells assessed by western blotting. ERK2 was used as a loading control. For gel source data, see Supplementary Fig. 1 (n = 3 experiments). j, Histogram of IL-15Rα measured by antibody-based staining and flow cytometry in MDA-MB-231, 4T07 and 4T1 cells. k, Quantification of the relative percentages of quiescent (Tomato+mVenus+) and cycling (Tomato+mVenus−) cancer cells after 24 h of treatment with IL-15 shows no effect on cell population ratios (n = 3 independent experiments). l–n, Flow cytometry quantification of T cell frequency (l) and activation (m, n) in livers from 4T1-injected mice treated with PBS or IL-15 (n = 5 PBS, n = 5 IL-15). In a, c, d, g, h, k–n, mean ± s.d.; two-tailed nonparametric Mann–Whitney U test.
