Fig. 3

Unsupervised clustering of T-cell subsets according to the level of membrane protein expression. Unsupervised clustering analysis of flow cytometric dataset using PhenoGraph algorithm (n = 34). A UMAP displaying the 25 clusters defined based on the fluorescence intensity of each marker tested, including ICPs. B Heatmap showing the protein expression patterns in each cluster. Fluorescence intensity of each marker has been normalized independently. C Pie charts representing the relative abundance (mean) of each cluster in the whole cohort, in CD8⁺, CD4⁺FoxP3⁻ and CD4⁺FoxP3.⁺ T cells (left panels); stacked bar chart displaying the relative abundance of each cluster in each tumor specimen in the 3 T-cell subsets (right panels). ICPs: immune checkpoints: (CD25, CD28, CD39, 4-1BB, CTLA-4, ICOS, OX40, PD-1, PD-L1, and TIGIT)