Fig. 1 |. PGE₂ EP2 receptor regulates myeloid metabolism and inflammation in ageing.

Data are mean ± s.e.m. unless otherwise specified. a, Levels of PGE₂ from young and aged human MDMs cultured for 20 h; ****P < 0.0001 by two-tailed Student’s t-test. b, Real-time changes in the OCR of human MDMs in response to treatment with the indicated concentrations of PGE₂ for 20 h. Cells were treated with 1 μM oligomycin (olig), 2 μM carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and 0.5 μM rotenone and antimycin (rot/an), as indicated by the three black arrows. Veh, vehicle. c, Basal respiration (OCR) (blue) and ECAR (red) in human MDMs stimulated with the indicated concentrations of PGE₂ for 20 h. Box plots (upper box limit, 5th percentile; lower box limit, 95th percentile; centre line, median; upper whisker, maximum value; lower whisker, minimum value); one-way analysis of variance (ANOVA), P < 0.0001; Tukey’s post hoc test, **P = 0.0085, ^†P = 0.0001, ^#P < 0.0001. d, Basal respiration and ECAR in human MDMs stimulated with the indicated concentrations of the EP2 agonist butaprost for 20 h. Box plots (5th–95th percentile); one-way ANOVA, P < 0.0001; Tukey’s post hoc test ^#P < 0.0001. In a–d, n = 5 donors per group; age (mean ± s.e.m.) 47.8 ± 2.105 years. e, Basal respiration and ECAR in peritoneal macrophages from young (3–4 months old) and aged (20–23 months old) Cd11bCre and Cd11bCre;EP2^lox/lox mice. Two-way ANOVA, age and genotype P < 0.0001; Tukey’s post hoc test, ****P < 0.0001 (n = 5 mice per group). f, TEM of peritoneal macrophage mitochondria from young and aged Cd11bCre (red arrows) and Cd11bCre;EP2^lox/lox (blue arrows) mice; two independent experiments (n = 6 mice per group). Scale bars, 100 nm. g, Quantification of TEM mitochondrial metrics from f. Box plots (5th–95th percentile); two-way ANOVA, age and genotype P < 0.0001; Tukey’s post hoc test, ****P < 0.0001 (n = 106 cells per group). h, Flow cytometry histograms of peritoneal macrophages for the anti-inflammatory markers CD71 and EGR2 and the pro-inflammatory markers CD80 and CD86 from young and aged Cd11bCre and Cd11bCre;EP2^lox/lox mice; three independent experiments (n = 10,000–20,000 cells per sample; n = 3 mice per group). i, Phagocytosis of fluorescent Escherichia coli particles in peritoneal macrophages from young and aged Cd11bCre and Cd11bCre;EP2^lox/lox mice. Two-way ANOVA, age P = 0.0147 and genotype P = 0.0008; Tukey’s post hoc test, *P = 0.0127, **P = 0.0017 (n = 6 mice per group). j, Hierarchical clustering of significantly regulated immune factors in the plasma and hippocampus from young and aged Cd11bCre and Cd11bCre;EP2^lox/lox mice (n = 10 young, n = 6 aged mice per group).