Extended Data Fig. 2: Cdk5 promotes CD8+ T cell infiltration in mouse and human BrM and boosts the antigen-specific CD8+ T cell killing in vitro. | Nature Cell Biology

Extended Data Fig. 2: Cdk5 promotes CD8+ T cell infiltration in mouse and human BrM and boosts the antigen-specific CD8+ T cell killing in vitro.

From: Astrocyte-induced Cdk5 expedites breast cancer brain metastasis by suppressing MHC-I expression to evade immune recognition

Extended Data Fig. 2

A. UMAP plots illustrate predicted immune populations in BrM from EO771.Br3.shCtl or EO771.Br3.shCdk5 cells (N = 3 mice per group). T cell clusters are circled in red. B. Major immune populations in BrM from EO771.Br3.shCtl or EO771.Br3.shCdk5 cells (N = 3 mice per group) based on scRNA-seq analysis. N = 4 technical replicates. C. Major T cell subsets in BrM from EO771.Br3.shCtl or EO771.Br3.shCdk5 cells (N = 3 mice per group) based on scRNA-seq analysis. N = 4 technical replicates. D. IHC profiling of major immune populations in BrM lesions of mice ICA-injected with EO771.Br3 or 4T1.Br3 cells transfected with control or Cdk5 shRNA (N = 3 mice per group). Cell number was normalized to BrM area. Each dot indicates a BrM lesion. Two-sided T-test. E. Representative IHC staining of CD8+ TILs in BrM lesions of mice ICA-injected with T11.Br1 cells transfected with control or Cdk5 shRNA (N = 3-5 mice per group). Scale bar = 100 μm. F. Two-sided Spearman’s correlation analysis of the expression of CDK5 and 3-gene TIL signature (CX3CR1, FGFBP2, FCGR3A) in 35 breast cancer BrM tissues from the GSE52604 human dataset. Each dot represents a patient. G. Spearman’s correlation analysis of the expression of CDK4/CDK6 and 12-gene TIL signature (CD8A, CCL2, CCL3, CCL4, CXCL9, CXCL10, ICOS, GZMK, IRF1, HLA.DMB, HLA.DOA, and HLA.DOB) in 35 breast cancer BrM tissues from the GSE52604 human dataset. Each dot represents a patient. H. Design of an experiment to test whether Cdk5 affects antigen-specific T cell killing of BrM cells in vitro. I. Quantification of remaining live cells 48 h after co-culture of indicated cancer cells with or without OT-I T cells (1:1 effector:target ratio). N = 3 biological replicates. Two-way ANOVA with Tukey’s post-hoc test. J. Representative images of culture wells from the experiment in J. Scale bar = 400 μm. Throughout, results are shown as means ± SEM. N.S. – not significant. TIL – tumour-infiltrating lymphocytes.

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