Fig. 1

PDIA3 inhibits mitochondrial respiratory function in cultured human brain endothelial cells. a PDIA3 gene deletion or PDIA3-/- (KO) was achieved in immortalized hCMEC/D3 cells (CMECs) by CRISPR-Cas9 and confirmed by RT-qPCR (delta–delta-Ct normalized first to GAPDH) and western blotting (b). WT CMECs were transfected with non-targeting gRNA. c Genotype was further confirmed by PCR from genomic CMEC DNA surrounding the target cut site. d Respiratory rates, as measured by oxygen consumption rate (OCR) by Seahorse analysis, were increased in PDIA3-/- CMECs relative to WT cells (repeated measures ANOVA. n = 3 per condition). e Reserve (spare) capacity (Res. Cap.) and maximal respiratory capacity (Max. Resp.) were increased but not basal respiratory rate and ATP-linked respiration (n = 3 each, two-way ANOVA). f The PDIA3-/- phenotype of increased bioenergetic function was largely reversed by overexpressing WT human PDIA3 compared to GFP control transfection (g)