Fig 1: Necroptosis promotes tumor metastasis by activating cell surface proteases. a, b ELISA of soluble JAM-A, CD138 and E-cad in TIF(a) and serum (b) from WT and Mlkl KO MMTV-PyMT mice at 15 weeks (n = 6, each). c, d ELISA of soluble JAM-A, CD138 and E-cad in TIF (c) and serum (d) from MMTV-PyMT mice treated with vehicle or GW280264X (n = 5, each). e qPCR analysis of IFN-γ mRNA expression in tumor-infiltrating CD8+ T cells of MMTV-PyMT mice treated with vehicle or GW280264X (n = 5, each; left panel). Flow cytometry analysis of tumor-infiltrating GZMB+ CD8+ T cells in MMTV-PyMT mice treated with vehicle or GW280264X (n = 5, each; right panel). f qPCR analysis of IFN-γ and T-bet mRNA expression in tumor-infiltrating CD4+ T cells of MMTV-PyMT mice treated with vehicle or GW280264X (n = 5, each). g Flow cytometry analysis of peripheral blood GZMB+ CD8+ T cells and IFN-γ+ CD4+ T cells MMTV-PyMT mice treated with vehicle or GW280264X (n = 5, each). h Left panel shows the representative images of H&E-stained lung sections from MMTV-PyMT mice treated with vehicle or GW280264X.Scale bar, 2 mm. Right panel shows the quantification of metastatic foci in lungs from MMTV-PyMT mice treated with vehicle or GW280264X (n = 5, each). Data presented as the mean ± sem and p value was determined by two-tailed t test (a–h)