Figure 7.

Metformin reduces hypoxia-dependent apoptosis in human CD8 T cells. (A, B) Viable cell count (A) and percentage of dead cells (B) of human CD8 T cells activated with anti-CD3/CD28 beads and feeder cells and incubated in normoxia (21% O₂) or hypoxia (1% O₂) and treated or not with metformin (2 mM). (C) Quantification of late apoptosis measured by fluorescence-activated cell sorting (FACS) staining for Annexin V and 7-aminoactinomycin D (7-AAD) in human CD8 T cells treated as in (A, B). (D, E) Lymphocyte-activation gene 3 (LAG3) (D) and programmed cell death protein 1 (PD-1) (E) expression measured by FACS in cells treated as in (A, B). Results are expressed as fold change compared with control. (A–E) is a pool of three independent experiments. All conditions are assessed in triplicate in each individual experiment. All data are mean±SEM. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001, calculated by two-tailed unpaired t-test. MFI, median fluorescence intensity.