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Fig. 3 | Journal for ImmunoTherapy of Cancer

Fig. 3

From: PD-1 signaling affects cristae morphology and leads to mitochondrial dysfunction in human CD8+ T lymphocytes

Fig. 3

PD-1 inhibits mitochondrial function in activated CD8+ T cells. a Representative dot plots of CD8+ T cells stained with MitoTracker Green and TMRM to determine the effect of the indicated stimuli on mitochondrial polarization. Incubation with the depolarizing agent DNP was used as negative control. b Time-dependent expansion of TMRM+ cells after indicated stimuli (n = 5). c Representative histograms of TCTRL, TACT and TACT + PD1 cells after 48 h stimulation. TMRM fluorescence is shown of DNP-treated TACT cells (negative control; dotted line). d Mean fluorescence intensity of TMRM+ CD8+ T cells at different times post-stimulation, assessed from data as in c (n = 5). e Percentage of ROS+ cells as detected with the MitoSOX Red probe. f Scheme for analyzing the reversibility of PD-1 effects on mitochondrial potential. g, h Percentage of TMRM+ cells and TMRM mean fluorescence intensity in TACT and pre-treated TACT + PD1 cells re-stimulated with TCTRLand TACT beads (n = 3). Data shown as mean ± SEM. *** p < 0.001, two-way ANOVA with Bonferroni post-hoc test; * p < 0.05, two-tailed paired Student’s t-test

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