Fig. 1

Enhanced antitumor immune responses in Rnf5^−/− mice. a Growth of YUMM1.5 (Braf^V600E:PTEN^−/−:Cdkn2a^−/−) melanoma cells after subcutaneous injection of 10⁶ cells into WT or Rnf5^−/− mice (n = 5). b Quantification of tumor-infiltrating effector (CD44^hi) CD4⁺ and CD8⁺ T cells and total CD45⁺ cells on day 24 after tumor injection (n = 5). c Frequencies of tumor-infiltrating TNF-α-, IFN-γ-, and IL-2-producing CD4⁺ and CD8⁺ T cells on day 24 after tumor inoculation (n = 5). d Quantification of tumor-infiltrating total DCs, pDCs, mDCs, and CD8α⁺ DCs on day 24 after tumor inoculation (n = 5). e Expression (mean fluorescence intensity, MFI) of MHC class II, CD40, CD80, and CD86 on tumor-infiltrating DCs (CD45⁺ CD11c⁺) on day 24 after tumor inoculation (n = 5). f Quantification of OT-I CD8⁺ T cells in tumor-draining lymph nodes (TdLN) and non-draining lymph nodes (ndLN) of CD45.1⁺ WT and Rnf5^−/− mice injected with B16-OVA melanoma cells (WT, n = 6; Rnf5^−/−, n = 5). g, h Growth of YUMM1.5 melanoma cells in mice injected i.p. with control IgG and anti-CD4 (g) or control IgG and anti-CD8 (h) depleting antibodies on days 0, 3, 6, 11, 16 (n = 9). FACS analysis revealed >90% depletion of blood CD4⁺ and CD8⁺ T cells on day 7 after tumor inoculation. i, Growth of YUMM1.5 melanoma cells in lethally irradiated bone marrow-reconstituted WT or Rnf5^−/− mice (arrow indicates bone marrow donor → recipient; n = 7). Data are representative of three independent experiments (a–e), two independent experiments (f, i) and one experiment (g, h) ≥5 mice per group. Graphs show the mean ± s.e.m. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001 by two-way ANOVA with Sidak’s correction (a, g–i) or by two-tailed t-test or Mann–Whitney U test (b–f)