Fig. 3: CD31 regulates EBF via its effects on EC metabolism. | Nature Communications

Fig. 3: CD31 regulates EBF via its effects on EC metabolism.

From: Preservation of microvascular barrier function requires CD31 receptor-induced metabolic reprogramming

Fig. 3

a EC were pre-treated with the glucose analog 2-DG (5 mM, 2 h) then MHC-1- or ICAM–stimulated for 4 h prior to TEER measurements. The bar graph shows the mean of measurement collected in three separate experiments of identical design ± SD. One-way Anova with Tuckey post-hoc test. 2DG MHC vs all ***p = 0.0003. b Glut-1 expression by EC following MHC stimulation (2 h) was evaluated by qRT-PCR. Data are mean ± SD. Two-tailed Student’s T test. MHC vs IsC *p = 0.0412. c, d Representative histograms of antibody-stimulated WT or CD31-deficient EC incubated with 6-NBDG for 2 h prior to analysis. 2-DG was used as a negative control. The graph shows the mean MFI measured in three independent experiments ± SD. One-way Anova with Tuckey post-hoc test. WT IsC vs WT MHC **p = 0.003, cd31−/− IsC vs cd31−/− MHC ns = 0.058. e ATP levels were measured in WT and CD31-deficient EC 4 h after MHC or insulin (1.8 µM) stimulation. Data are mean ± SD. Two-tailed Student’s T test. WT MHC vs cd31−/− MHC ***p = 0.0002, WT insulin vs cd31−/− insulin **p = 0.0062. Extracellular acidification rate (ECAR) of unstimulated and antibody-stimulated WT and CD31-deficient EC is shown in (f–h). The basal and maximal glycolysis and the glycolytic reserve are shown in (i–k). Wells were first injected with anti MHC antibodies at the indicated time point. Isotype-matched and secondary antibodies were used as controls. Further injections followed at the time point indicated (arrows) introducing the indicated compounds into the wells. N = 3 independent experiments. The error bars represent SD. One-way Anova with Tuckey post-hoc test. f WT time 81 vs cd31−/− time 81 **p = 0.0023, WT time 94 vs cd31−/− time 94 ***p = 0.0017; h IsC Glu/Oligo injection vs MHC Glu/Oligo **p = 0.012, IsC Oligo/2DG vs MHC Oligo/2DG *p = 0.027; i WT MHC vs WT IsC ****p < 0.0001, WT IsC vs cd31−/− IsC/MHC *p = 0.0245, WT MHC vs cd31−/− IsC/MHC ****p < 0.0001; j WT IsC vs cd31−/− IsC **p = 0.0056, WT IsC vs cd31−/− MHC **p = 0.0097, WT MHC vs cd31−/− IsC **p = 0.0024, WT MHC vs cd31−/− MHC **p = 0.0041; k WT IsC vs cd31−/− IsC ***p = 0.0002, WT IsC vs cd31−/− MHC ***p = 0.0003, WT MHC vs cd31−/− IsC ***p = 0.00024, WT MHC vs cd31−/− MHC ***p = 0.00021.

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