Figure 4 from Extracellular Vesicle Secretion by Leukemia Cells in Vivo Promotes CLL Progression by Hampering Antitumor T-cell Responses

LME-sEVs impact CD8+ T-cell transcriptome, proteome, and metabolome. A, Volcano plot showing DEG identified by RNA-seq from CD8+ T cells treated for 48 hours with LME- (n = 4) or HCME-sEVs (n = 3) with FDR <0.05 and log2FC >1. B and C, Hierarchical clustering of all DEG (B) and of selected genes from relevant cell functions (C). D, Volcano plot showing differentially expressed proteins (DEP) identified by mass spectrometry from CD8+ T cells treated for 96 hours with LME- (n = 3) and HCME-sEVs (n = 3) with FDR <0.05 and log2FC >1. E and F, GzmB mRNA expression and GzmB and perforin levels in CD8+ T cells treated for 48 hours with HCME- or LME-sEVs. G and H, Ontology analysis of enriched (G) or diminished (H) DEP in CD8+ T cells treated with LME- or HCME-sEVs (from D). I, Levels of glucose measured by mass spectrometry in culture medium in CD8+ T-cell treated with LME- or HCME-sEVs for 96 hours. Negative value represents consumption. J, Immunoblot analysis of glycolysis-related proteins from CD8+ T cells treated for 96 hours with LME- or HCME-sEVs. K, Levels of ADP and ATP generated from 13C-glucose measured by mass spectrometry in CD8+ T cells treated with LME- (n = 5) or HCME-sEVs (n = 6) for 96 hours. L, Oxygen consumption measured by SeaHorse assay from CD8+ T cells treated with LME- or HCME-sEVs for 96h. *, P < 0.05; **, P < 0.01; ****, P < 0.0001 (unpaired Student t test). Data are mean and SEM.