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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Immuno-metabolic impact of the multiple sclerosis patients’ sera on endothelial cells of the blood-brain barrier

Fig. 4

RRMS sera induce impaired engagement of glycolysis in ECs from the BBB. a Kinetic profile of ECAR in hCMEC/D3 cells treated overnight with 20% HD or RRMS naïve-to-treatment sera. Real time ECAR was measured, under basal conditions and in response to glucose, oligomycin and 2-DG. Indication of glycolytic pathway activation, calculated from hCMEC/D3 ECAR profile: (b) basal glycolysis, (c) glycolytic capacity and (d) glycolytic reserve. Data are presented as average of three experiments performed on 25 donors/group, matched in sex, age and EDSS score. Data are shown as mean ± s.e.m. Statistical analysis was performed by paired Wilcoxon test **P ≤ 0.01, ***P ≤ 0.001). e, f Sera isolated from RRMS patients downregulate GLUT1 expression on membrane of brain ECs. hCMEC/D3 monolayer plated on 24-well plate were treated (16 h) with sera isolated from RRMS patients and matched HD. Cells were collected and stained with rabbit anti-GLUT-1 antibody. DAPI was used for nuclear staining. e Imaging flow cytometry was performed on an ImageStreamx Mark II operated by INSPIRE software (Amnis Corporation). f Data represent % of mean intensity of fluorescence (MIF) in comparison to sera from matched HD. Statistical analysis was performed using Mann-Whitney U test (two-tails) *P < 0.05 vs HD, n = 5/group

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