Fig. 5: The metabolic response to endothelial barrier breach is src-phosphatase-dependent. | Nature Communications

Fig. 5: The metabolic response to endothelial barrier breach is src-phosphatase-dependent.

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

Fig. 5

Confluent EC monolayers were MHC-stimulated and treated with the selective SHP1/2 inhibitor (50 μM for 3 h) or vehicle as a control. a The bar graph shows the mean TEER values measured in three independent experiments. b Percentage of EC viability following treatment with SHP1/2 inhibitor determined by the trypan blue exclusion assay in three independent experiments. c–f ECAR measured in WT (c, d) or cd31−/− EC (e, f) following MHC antibody-stimulation with or without pre-treatment with the SHP1/2 inhibitor (50μM for 45 min). Basal and maximal glycolysis and glycolytic reserve are shown in (d, f). g–j OCR measured in WT g–h or CD31-deficient EC I, j following MHC antibody-stimulation with or without pre-treatment with the SHP1/2 inhibitor. Basal and maximal respiration and spare capacity are shown in (i, j). (N = 3 independent experiments). Data are shown as mean ± SEM; significant differences were determined using one-way Anova with Tuckey post-hoc test. a IsC vehicle vs MHC vehicle ****p < 0.0001, MHC vehicle vs MHC + SHP inhibitor ****p < 0.0001; c Glu/Oligo injection WT MHC vs all ****p < 0.0001, Oligo/2DG injection WT MHC vs all *p = 0.0103; d basal glycolysis WT IsC vs WT MHC **p = 0.0039, Basal glycolysis WT MHC vs WT + SHP inhibitor *p = 0.047, Glycolytic reserve WT MHC VS WT + SHP inhibitor *p = 0.0482, Max glycolysis WT IsC vs WT MHC ***p = 0.0008, Max glycolysis WT MHC VS WT + SHP inhibitor **p = 0.0015; e Glu/Oligo/2DG injection cd31−/− MHC + SHP Inhibitor vs all ****p < 0.0001; f basal glycolysis cd31−/− MHC vs cd31−/− MHC + SHP inhibitor **p = 0.019, glycolysis reserve cd31−/− MHC vs cd31−/− MHC + SHP inhibitor ***p = 0.0003, max glycolysis cd31−/− MHC vs cd31−/− MHC + SHP inhibitor *0.0384; i FCCP/Rotenone injection cd31−/− MHC + SHP inhibitor vs all ****p < 0.0001, FCCP/Rotenone injection cd31−/− Isc vs cd31−/− IsC+SHP inhibitor ***p = 0.0006; j max respiration cd31−/− IsC vs cd31−/− MHC *p = 0.0177, Max respiration cd31−/− IsC vs cd31−/− MHC + SHP inhibitor ***p = 0.0004, spare capacity cd31−/− IsC vs cd31−/− MHC ****p < 0.0001, cd31−/− IsC vs cd31−/− MHC + SHP inhibitor ****p < 0.0001.

Back to article page