Fig. 2: NK-specific gp96 deficiency reduces NK cell anti-tumor functions.
Female WT and KO mice were subcutaneously inoculated with LLC (A) or MC38 (B) tumor cells. Tumor size was measured every 2 days. C Endpoint tumor mass in mice 30d after MC38 cell inoculation. T-distributed stochastic neighbor embedding (tSNE) plots show the distribution of lymphocyte clusters in the spleen (D) and tumor tissue (E) as determined by flow cytometry analysis. F The absolute number of tumor-infiltrating NK cells from KO and WT mice as in (E). G Flow cytometry analysis of the frequency of cells expressing CD107a among tumor-infiltrating NK cells from MC38-bearing mice. H Representative lung nodules from WT and KO mice 2 weeks after intravenous (i.v.) injection of 0.25 × 106 B16F10 melanoma cells. I Hematoxylin and eosin (H&E) staining of lung sections from mice as in (H). Bar, 100 μm. J Frequency of cells expressing CD107a or IFN-γ among tumor-infiltrating NK cells from mice as in (H), analyzed 14d after challenge. K Assessment of natural cytotoxicity of FACS-sorted splenic NK cells against YAC-1 target cells labeled with CFSE. L Frequency and the absolute number of remaining CFSE-labeled YAC-1 cells in the peritoneal cavities of KO and WT mice. CFSE-labeled YAC-1 cells were intraperitoneally injected and evaluated 24 h post-injection. The data are representative of two independent experiments with similar results. Dots represent data from n = 5 mice/group (A–C, F–J, K) and n = 3 mice/group (D, E). Mean ± SD is shown. Statistical significance was determined using two-tailed unpaired t test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. p values: (C) p = 0.0106, (D) p = 0.0478 (macrophage), p = 0.0114 (NK), (E) p = 0.0016 (DC), p = 0.0275 (NK), (F) p = 0.005, (G) p = 0.0004, (H) p < 0.0001, (J) p = 0.0359 (CD107a+), p = 0.0031 (IFN-γ), (K) p = 0.0236 (5:1), p = 0.0065 (10:1), (L) p < 0.0001.
