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. 2021 Feb 10;9(2):e002044. doi: 10.1136/jitc-2020-002044

Figure 6.

Figure 6

Targeting transforming growth factor-β (TGFβ) restores the function and metabolism of natural killer (NK) cells from patients with breast cancer. Peripheral blood mononuclear cells from patients with breast cancer were stimulated with interleukin (IL)2 (500 IU/mL) (A–D), or with IL12 (30 ng/mL) and IL15 (100 ng/mL) (B, E, F) for 18 hours in the presence of isotype control or anti-TGFβ antibody (5 µg/mL). (A–C) Patient NK cells were stained for CD25, pS6 and CD71 and analyzed by flow cytometry. (D) Patient NK cells were stained with MitoTracker Green (100 nM) and analyzed by flow cytometry. (E) Representative oxygen consumption rate (OCR) trace. Inhibitors added at times indicated were oligomycin (Oligo), carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone (FCCP) and antimycin-A/rotenone (AntiA/Rot). (F) Pooled data for basal oxidative phosphorylation (OXPHOS) and maximal respiration. (G) Ratio of OCR:extracellular acidification rate (ECAR) in patient NK cells. (H) Representative dot plot for interferon-γ (IFNγ) production and pooled data for IFNγ production. Individual donors are shown by a dot (n=8–14). Samples were compared using a paired Student’s t-test, *p<0.05, **p<0.01.