Fig. 4: Orthogonal characterisation of a CD3, CD4 and CD8 immune-cold CRC phenotype.

PCA (a) and heatmap (b) demonstrating the distribution and clustering of the 20 most variable probes identified by differential gene expression analysis between immune-cold (Group A) and immune-NOS (Group B) subgroups in the S:CORT FOCUS cohort. GSEA enrichment plot (c) for the WINTER_HYPOXIA_METAGENE signature in the immune subgroups. Kaplan–Meier curve of dichotomised Winters Hypoxia Metagene Signature (d). Boxplot to demonstrate the relationship between immune subgroups and tumour hypoxia using CAIX IHC expression from the tumour epithelium (e). Kaplan–Meier curve of combined immune subgroups and tumour hypoxia (f). Differences in immune subgroups were compared using ANOVA. Differences in survival curves are presented as log-rank P value. Immune cold = patient stratification by collective low-density cell counts for CD3, CD4 and CD8 IHC; immune–not otherwise specified (NOS) = any other combination of CD3, CD4 and CD8 IHC expression; in e, f this was further stratified by hypoxia-low = low tumour hypoxia by the Winters Hypoxia Metagene Signature or hypoxia-high = high tumour hypoxia using the Winters Hypoxia Metagene Signature.