Skip to main content
. 2021 Aug 16;10:e68484. doi: 10.7554/eLife.68484

Figure 1. Hypoxia-inducible factor-1α (HIF1α) expression in natural killer (NK) cells.

(A) HIF1α protein expression in splenic NK cells (black) ex vivo compared to FMO control (gray) by flow cytometry. Representative experiment of two independent experiments. (B) Comparison of lymphocytes HIF1α protein expression. Bar graphs show data from two independent experiments with four mice per group. (C) Spleens from C57BL/6 mice were prepared as in Materials and methods and were stimulated with IFNβ (200 units) + IL-18 (50 ηg/ml) for 24 hr in normoxia (N, 20%) or hypoxia (H, 1%). HIF1α expression in NK cells was determined. Data are from two independent experiments with three mice per group. (D) Splenic NK cells were cultured in 25 ηg/ml IL-15 for 48 hr and HIF1α expression was measured at 16, 24, and 48 hr. Data are from two independent experiments with four mice per group. (E) CD69 and HIF1α was determined at day 1.5 post-infection (pi). Data are from 2–3 independent experiments with three mice per group. Statistical significance for uninfected versus indicated dose. (F) HIF1α expression in Ly49H+ or Ly49H- NK cells over 7 days from C57BL/6 mice that were infected with 50K plaque forming unit (pfu) murine cytomegalovirus (MCMV). Statistical significance for d0 versus indicated day. Data are from two independent experiments with 3–7 mice per group. All data depict mean ± SEM, with each data set containing data indicated number of mice per group from independent experiments. Unpaired t-test was performed (A, B, D–F) or one-way ANOVA (C). Statistical significance indicated by n.s., no significant difference; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Figure 1.

Figure 1—figure supplement 1. Expression of hypoxia-inducible factor-1α (HIF1α) in natural killer (NK) cells.

Figure 1—figure supplement 1.

(A) Representative histogram of HIF1α expression in splenic and liver NK cells direct ex vivo. Data are from two independent experiments from five mice per group. (B) Splenocytes from C57BL/6 mice stimulated for 6 hr with either anti-NK1.1, PMA and ionomycin (PMA/Io), or no receptor stimulation (No Rec). For 24 hr culture, splenic NK cells were stimulated with anti-NK1.1, or no receptor stimulation (No Rec) in the presence of low IL-15. IFNγ (left) and HIF1α (right) expression was measured in splenic NK cells. Data are from 2–3 independent experiments with 3–5 mice per group. (C) CD69 expression of splenic NK cells 3 days post murine cytomegalovirus (MCMV) infection, and HIF1α expression in CD69- and CD69+ NK cells (left) with quantification (right). Data are from three independent experiments with six mice per group. Data depict mean ± SEM, with each data set containing data indicated number of mice per group from independent experiments. Statistical significance indicated by n.s., no significant difference, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.