Supplementary Figure 4: Inhibitory and exhaustion-associated TIM-3 is increased on VHL-deficient cells and reduced PD-1 expression on VHL-deficient cells is HIF-1α-HIF-2α independent and VHL-dependent.
From: Hypoxia-inducible factors enhance the effector responses of CD8+ T cells to persistent antigen
(a) Cotransfer of VHL-sufficient and VHL-deficient P14 CD8+ T cells followed by LCMV clone 13 infection as in Fig. 2a; histograms of TIM-3 expression and gMFIs of P14 CD8+ T cells from spleen on day 17 of infection with LCMV clone 13; n=4, * P = 0.008 (Student's unpaired t-test). (b) Cotransfer of VHL-sufficient and VHL-deficient P14 CD8+ T cells followed by LCMV clone 13 infection as in Fig. 2a; flow cytometric analysis of splenic VHL-sufficient P14 (black) or VHL-deficient P14 (red) CD8+ T cells at the indicated time points, gMFI shown below. Expression of PD-1 on total host CD8α+ T cells excluding P14 donor cells (histograms, grey), or gMFI for PD-1 low –expressing cells (in graphs, open bars) shown for reference; n=3, * P = 0.004, ** P = 0.05 and *** P = 0.004 (Student's unpaired t-test). (c) Experimental design for a mixed transfer of VHL-sufficient, VHL-deficient and VHL-HIF-1α-HIF-2α-triple-deficient P14 CD8+ donor cells. (d) Histogram of each donor population displaying PD-1 (left) and gMFI graphed (right); peripheral blood on day 7 of infection; n=3, error bars indicate s.e.m.
