Fig. 3

Identification of CD4⁺ T-cell clusters in Tumor Infiltrating T-cell populations (a) Heatmap of all CD4⁺ T-cell clusters identified at day 8 after the start of the PD-L1 treatment. Data shown is based on t-SNE plots, and is pooled from the control and PD-L1 treated group. Level of ArcSinh5-transformed expression marker is displayed by a rainbow scale. Dendrogram on the top represents the hierarchical similarity between the identified clusters. (b) Average and SEM in percentage of each CD4⁺ T-cell cluster among the CD4⁺ T-cell population of control (blue bars) and PD-L1 group (red bars). (c) t-SNE plot of respectively 0.23 × 10⁶ and 0.25 × 10⁶ CD4⁺ T cells from control (blue) and PD-L1 treated (red) group. (d) Same t-SNE plots as above now showing the level of expression marker by a rainbow scale. The arrow identifies the cluster of interest 12 (having a shared CD4⁺ LAG3⁺ ICOS⁺ phenotype)

(e) Bar graph showing the mean frequency of cluster 12 (± SEM, unpaired t-test). Individual mice belonging to the control (blue) and PD-L1 treated (red) group are indicated.