Fig. 2. Dapl1 deficiency promotes Tim3 expression and improves responses to ICR blockade.

a,b, Flow cytometry analysis of the frequency of Tim3⁺, PD1⁺ and Lag3⁺ CD8 T cells (a), and the frequency and absolute cell number of Tim3⁺ PD1⁺ CD8 T_EX cells (b) in the tumor of B16-OVA-implanted wildtype (WT) and Dapl1 KO mice. n = 5 per genotype. c, Flow cytometry analysis of the frequency of TCF1⁺Tim3⁻, TCF1⁺Tim3⁺ and TCF1⁻Tim3⁺ subsets, gated on CD8⁺PD1⁺ T cells in the tumor of B16-OVA tumor-bearing wildtype or Dapl1 KO mice, n = 5 per genotype. d,e, Flow cytometry analysis of the frequency of Tim3⁺ and PD1⁺ CD8 T cells (d), TCF1⁺Tim3⁻, TCF1⁺Tim3⁺ and TCF1⁻Tim3⁺ subsets(e) in the tumor of B16F10-implanted B6.SJL mice (CD45.1⁺) adoptively transferred with Dapl1 KO Pmel1 CD8 T cells (CD45.2⁺) transduced with either an empty vector (Mock) or HA-Dapl1 expression vector (Dapl1). n = 5 per genotype. f-h, Tumor growth curves (f), survival curves (g) and flow cytometry analysis of the frequency of IFN-γ- or TNF-α- producing CD8 T cells in the tumor (h) of wildtype or Dapl1 KO mice injected subcutaneously with B16-OVA followed by i.p. injection with anti-PD1 (100 μg/mouse) or isotype control on days 7, 9, and 11. n = 6 (f,g) or n = 5 (h) per genotype. i,j, Tumor growth curves (i) and survival curves (j) of wildtype or Dapl1 KO mice injected subcutaneously with B16-OVA followed by i.p. injection with anti-Tim3 (200 μg/mouse) or isotype control on days 7, 9, and 11. n = 7 per genotype. k, Flow cytometry analysis of the frequency of TCF1⁺Tim3⁻, TCF1⁺Tim3⁺ and TCF1⁻Tim3⁺ subsets, gated on H2Db/GP_33–41 tetramer⁺ CD8⁺PD1⁺ T cells in the spleen of wildtype or Dapl1 KO mice infected with LCMV clone 13 for 30 days, n = 5 per genotype. Data are representative of three independent experiments. Summary data are shown as the mean ± s.d. with P values determined using a two-tailed unpaired Student’s t-test (a-e, h, k), two-sided log-rank Mantel–Cox tests (g, j) and two-way ANOVA with Bonferroni correction (f, i). ns, not significant.