Fig. 7.
PRE-084 fails to enhance endothelial glycolytic rate after siRNA-mediated knockdown of σ1. At 70 h post-transfection, confluent HUVEC monolayers transfected with SIGMAR1 siRNA (σ1 kd) were treated with PRE-084 or vehicle for 3 h. A. Time course of glycolytic proton efflux rate (GlycoPER) of σ1 kd cells versus NT control cells during the glycolytic rate assay, in which cells were treated with a rotenone/antimycin-A cocktail (Rot/AA) and later with 2-deoxyglucose (2-DG). B. Oxygen consumption rate (OCR) during the same assay. Additional parameters were calculated from the assay for basal glycolysis (C), compensatory glycolysis (D), basal protein efflux rate (E), the ratio of mitochondrial oxygen rate (mitoOCR) to GlycoPER, and the post-2-deoxyglucose acidification (G). The two groups were compared with unpaired t-tests for each parameter. P values are shown for all comparisons. N=7–9 HUVEC monolayers per group.