electric cell substrate impedance sensing  (Applied BioPhysics)

Journal: bioRxiv

Article Title: Mitochondrial carbonic anhydrase-VB inhibition rescues brain endothelial stress and memory in Alzheimer’s disease models

doi: 10.64898/2026.03.16.711716

Figure Lengend Snippet: A) hCMEC barrier resistance was assessed by ECIS Zθ to evaluate barrier integrity. Human CMEC were treated with 10µM Aβ40-Q22 (Q22), alone or in combination with 10 or 30µM 4ITP. B-C) Western blot analysis of occludin and oligomeric claudin-5 (normalized to actin, and plotted as % change of Ctrl), following 24h (B) and 48h (C) treatment. Data represents 3 individual experiments with 2 replicates per group, graphed as mean ± SEM. Statistical significance was evaluated by One-way ANOVA followed by Tukey post-hoc test. *P< 0.05, **P<0.01, ***P<0.001, ****P<0.0001

Article Snippet: Trans-endothelial electrical resistance was measured using the ECIS ZΘ system (Applied Biophysics).

Techniques: Western Blot

Journal: bioRxiv

Article Title: Mitochondrial carbonic anhydrase-VB inhibition rescues brain endothelial stress and memory in Alzheimer’s disease models

doi: 10.64898/2026.03.16.711716

Figure Lengend Snippet: CA-VB KO in hCMEC protects from Aβ-induced apoptosis and BBB permeability . A) WB confirmed CA-VB absence in KO cells. B) DNA fragmentation, plotted as fold of change (FOC) of untreated control (ctrl) cells, measured by cell death ELISA. C) Representative IF images of active caspase-3/7 (green) and quantification to the right in CMEC treated with 25µM Q22 for 24 hours. Original magnification: 10x . D) IF of CA-VB KO vs WT hCMEC to detect mitochondrial membrane potential (Mito-tracker, red) as well as cytochrome C (Cyto C, green), in hCMEC challenged with Q22 for 16 hours. Zoom images of the merge signal at the bottom depict altered mitochondrial network with perinuclear mitochondria in Q22-treated hCMEC, but not in CA-VB KO cells. Original magnification:100x E) Mitochondrial H 2 O 2 measured with Amplex Red kit. F) Barrier resistance was measured over time with ECIS-Zθ in WT and CA-VB KO hCMEC treated or not with 10µM Q22. Data represents 3 individual experiments with 2 replicates each, graphed as mean ± SEM. Statistical significance was evaluated by Two-way ANOVA followed by Tukey post-hoc test. *P< 0.05, **P<0.01 ****P<0.0001

Article Snippet: Trans-endothelial electrical resistance was measured using the ECIS ZΘ system (Applied Biophysics).

Techniques: Permeability, Control, Enzyme-linked Immunosorbent Assay, Membrane

Journal: Frontiers in Immunology

Article Title: SARS−CoV−2 spike S1-mediated HIF−2α activation in retinal endothelial cells suggests a mechanism contributing to post−COVID endothelial dysfunction

doi: 10.3389/fimmu.2026.1770758

Figure Lengend Snippet: S1 and plasma from PCS patients influence ROS production, impair NO availability, and disrupt barrier integrity in HRECs, effects improved by belzutifan. (A) HRECs were mock-treated (control) or stimulated with S1 (100 ng/ml) for 0–6 h, and cellular ROS levels were measured using DCFDA/H 2 DCFDA (n = 3 independent experiments). (B) HRECs were mock-treated (control) or stimulated with S1 (100 ng/ml) for 4 h, and mitochondrial ROS production was measured by flow cytometric analysis using MitoSox Red (n = 4 independent experiments). (C, D) HRECs were mock-treated (control), stimulated with S1 (100 ng/mL) or CoCl 2 (100 µM), and treated with belzutifan (50 nM) for 72 h. Immunofluorescence staining was performed for F-actin (C, red ) and VE-cadherin (D, green ) , with nuclei counterstained with DAPI (blue). Images (left) were acquired at 20× magnification, and scale bars represent 100 µm. Graphs (right) illustrate the percentage of positive cells (C) and the corrected total cell fluorescence (CTCF) (D) (n = 3 independent experiments). (E) HRECs were cultured at confluence on ECIS electrodes and then stimulated with 100 ng/mL S1 or left untreated in the presence or absence of 50 nM belzutifan for 0–72 h. The loss of barrier integrity was determined by transendothelial electrical resistance (TEER). Values were normalized to time = 0 for easier comparisons (n = 3 independent experiments). (F) HRECs were treated with 2% plasma from healthy individuals (HC, n=8) or PCS patients (n=13) for 0–6 h, and cellular ROS levels were measured using DCFDA/H 2 DCFDA. (G) Mitochondrial ROS production in HRECs exposed to 2% plasma from HC (n=8) or PCS patients (n=13) for 4 h, measured by flow cytometric analysis using MitoSox Red. (H) Total NO levels in HRECs exposed to 2% plasma from HC (n=8) or PCS patients (n=13) for 4 h and 24 h, measured using a fluorometric assay for total nitrite/nitrate levels. (I) HRECs were cultured at confluence on ECIS electrodes and exposed to 2% plasma from HC or PCS patients in the presence or absence of 50 nM belzutifan for 0–48 h. The loss of barrier integrity was determined by transendothelial electrical resistance (TEER). Values were normalized to time = 0 for easier comparisons. Data are represented as means ± SD. Each dot represents one independent experiment for S1 studies or one individual donor for plasma studies. A p-value of <0.05 was considered statistically significant. P-values were determined by two-way ANOVA followed by Tukey’s post hoc test (A, E, F, I) , Mann–Whitney U test (B) , one-way ANOVA followed by Tukey’s post hoc test (C, D) , Student’s t-test (G) , and Kruskal–Wallis test followed by Dunn’s post hoc test (H) . .

Article Snippet: The barrier function of confluent endothelial cell monolayers was estimated using electric cell‐substrate impedance sensing (ECIS) model Z-Theta (Applied Biophysics) as described ( ).

Techniques: Clinical Proteomics, Control, Immunofluorescence, Staining, Fluorescence, Cell Culture, MANN-WHITNEY