Extended Data Fig. 4: Characterisation of T cell responses and metastasis load in Arhgef1-deficient animals.

a, PD-1, TIM-3 and TIGIT expression on effector CD4⁺ T cells within tumour-bearing lungs of Cd4^Cre (cWT) and Arhgef1^fl/fl Cd4^Cre (Arhgef1 cKO) animals. Flow cytometry plots (left) show CD4⁺ T cells from tumour-bearing lungs, and graphs show replicate measurements from tumour-bearing (cWT = 11; Arhgef1 cKO = 10) and non-bearing (cWT = 4; Arhgef1 cKO = 4) animals (right). b, Intracellular cytokine staining of OVA_257-264-specific CD8⁺ T cells from tumor-bearing lungs of Cd4^Cre (cWT = 11) and Arhgef1^fl/fl Cd4^Cre (Arhgef1 cKO = 10) animals 17 days after intravenous (i.v.) injection of B78ChOva melanoma cells. Flow cytometry plots of TNF, IL-2 and IFN-γ expression (left) and frequency of cells expressing indicated cytokines upon brief stimulation ex vivo, presented in terms of cytokine polyfunctionality (right). c, Kinetic analysis of mCherry⁺ CD45⁻ melanoma cells within lungs from WT (n = 6-8) and Arhgef1 KO mice (n = 4-8) at the indicated time points after i.v. injection of B78ChOva cells. d, Relationship between tumour burden and PD-1 expression by CD44⁺ CD8⁺ T cells of lungs of Cd4^Cre (cWT = 17) and Arhgef1^fl/fl Cd4^Cre (Arhgef1 cKO = 18) animals 17 days after i.v. injection of tumour cells. Data are representative of two (a-c) or pooled from two (d) independent experiments. One-way ANOVA with Tukey multiple comparisons test (a); unpaired two-tailed Student t tests with Holm-Šídák correction for multiple hypothesis testing (b, c) and Simple Linear Regression (d). Graphs show mean ± s.e.m.

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