Fig. 2: Plck-GAPDH malignant CD4+ PD-1^high cells are addicted to mitochondrial respiration.

A FACS analysis of Tfh markers PD-1 and CXCR5 on CD4+ T cells in plck-GAPDH tumors compared to WT CD4+ splenocytes. B The percentage CD4+ PD-1^high cells in plck-GAPDH tumors compared to WT splenocytes per total CD4+ cells (mean ± SD, WT n = 12, plck-GAPDH, n = 27, ****p < 0.0001) and the number of CD4+ PD-1^high cells in plck-GAPDH lymphoma-bearing spleens and in WT spleens (mean ± SD, WT n = 12, plck-GAPDH, n = 27, ****p < 0.0001). C WT CD4+ splenocytes and plck-GAPDH tumor CD4+ and CD8+ T cells and CD19 + B cells were sorted by negative selection; the CD4+ tumor cells were further sorted for the PD-1^high fraction and PD-1 negative fraction). In each of these cell fractions we determined their mitochondrial and glycolytic metabolic ATP requirements using a method based on Cell Titer Glo Kit (mean is shown for n = 3; **p < 0.01, ****p < 0.0001). D, E PD-1, CD4, CD8, and CD19 surface antibody staining for WT splenocytes and plck-GAPDH tumors was followed by SCENITH metabolic FACS analysis to determine the dependance on glucose or mitochondria for the different cell subpopulations (E) and their glycolytic or fatty acid oxidation capacity (E) (mean ± SD, n = 3; **p < 0.01, ns not significant).