Fig 1: TIM3+ MICs induce an immunosuppressive environment during micrometastasis(A) Single cell RNA-seq uniform manifold approximation and projection (UMAP) of CD45+ immune cells isolated from liver metastasis after 4T07 Ctrl and Tim3-KD i.c. injection. UMAP representation of: Ctrl (red) and Tim3-KD (blue) in healthy liver adjacent tissue (AT), liver micrometastasis and liver macrometastasis.(B) scRNA-seq bubble plot showing average expression (color) and percentage expression (size) of different genes defining lymphoid annotated cell compartments across metastatic conditions. Average expression legend is shared among populations. Percentage expression is relative within each immune population type.(C) Idem B for the myeloid compartment.(D) scRNA-seq cell fraction of the lymphoid cell compartment in micrometastatic samples from 4T07 Ctrl and Tim3-KD conditions.(E) Flow cytometry analysis of CD4+ T cells, CD8+ T cells and γδ T cells from liver micrometastasis from 4T07-Ctrl and 4T07-Tim3-KD. Data represents mean ± SEM, n = 11 independent biological replicates in (E), (F), and (G).(F) Flow cytometry of cytotoxic γδ T cells (GZMB+) and immunosuppressive (IL17+) γδ T cells. Dot plot represents fold change of IL-17 and GZMB γδ T cell populations in TIM3+ (Ctrl) vs. TIM3- (Tim3-KD) micrometastasis.(G) Flow cytometry of cytotoxic CD8+ T cells (GZMB+, CD69+) and exhausted T cells (PD1+) populations. Dot plot represents fold change of PD1, CD69, and GZMB positivity in TIM3+ (Ctrl) vs. TIM3- (Tim3-KD) micrometastasis. Statistical significance respect to the Ctrl; ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001, by unpaired Student’s t test in (E), (F), and (G).Also see Figures S5, S6, and Table S2.