Journal: bioRxiv

Article Title: Urban PM 2.5 at Realistic Environmental Concentrations Impairs Blood–Brain Barrier Integrity and Enhances LOX-1 Expression in Human Brain Endothelial Cells

doi: 10.64898/2026.01.29.702473

Figure Lengend Snippet: A . Physiological rationale: Ambient PM2.5 exposure is epidemiologically linked to increased ischemic stroke risk. This in vitro model simulates the real-life scenario of pre-existing PM2.5 exposure followed by ischemic stroke and subsequent reperfusion. B . Primary adult male HBMEC were exposed to 5, 15, 75, or 300 μg/m 3 PM 2.5 for 48h in total. To compare with the effects of physiological ischemic-like injury, some plates were exposed to hypoxia (1% O 2 ) and glucose deprived media (HGD) for 3h after the initial 24h incubation. Following HGD or normoxia, cells were reperfused with nutrient-enriched media and incubated with PM 2.5 at normoxic (21% O 2 ) conditions as a reference for resolution of ischemia. Barrier integrity, cell viability, reactive oxygen species (ROS), inflammation and LOX-1 expression was assessed. Figure created in BioRender.

Article Snippet: Primary adult male HBMEC were purchased from Innoprot (Spain, Catalog number: P10361, Lot number: 111224CS).

Techniques: In Vitro, Incubation, Expressing

Journal: bioRxiv

Article Title: Urban PM 2.5 at Realistic Environmental Concentrations Impairs Blood–Brain Barrier Integrity and Enhances LOX-1 Expression in Human Brain Endothelial Cells

doi: 10.64898/2026.01.29.702473

Figure Lengend Snippet: Adult male HBMEC were exposed to vehicle or PM 2.5 (5, 15, 75, or 300 μg/m 3 ) for 24h and incubated for 3h in normoxia- or hypoxia and glucose deprivation (HGD) followed by 24h reperfusion. A . Live cell count (CyQUANT nuclear stain) decreased when exposed to ≥75 μg/m 3 PM 2.5 compared to vehicle. HGD treatment reduced live cell count compared to normoxia but did not differ between particle treated groups. B . Reactive oxygen species (ROS) signal (DCHF-DA) normalized to live cell count. Relative ROS levels increased dose-dependently with PM 2.5 concentration, with significant increase observed at PM 2.5 ≥75 μg/m 3 , in comparison to normoxia vehicle. ROS levels were uniformly elevated following HGD across all doses in comparison to normoxia vehicle and significantly higher than untreated HBMEC. (n=12 technical replicates for vehicle and 5, n=8 technical replicates for 15, 75 and 300) C . Analysis of crystal violet-stained HBMEC shows a longer maximum cellular length when treated with ≥15 μg/m 3 PM 2.5 . (n=21-37 individual cells) D . Representative images of crystal violet-stained HBMEC visualizing a differentiated morphology in cells treated with higher PM 2.5 concentration, where cells appear more elongated and expanding towards neighbouring cells. Data presented as mean ± SD. Statistical significance assessed through Kruskal-Wallis test within treatment groups (Normoxia/HGD) and Mann-Whitney test between groups with different treatment (300 normoxia/vehicle HGD). *p<0.05. ***p<0.001. ****p<0.0001.

Article Snippet: Primary adult male HBMEC were purchased from Innoprot (Spain, Catalog number: P10361, Lot number: 111224CS).

Techniques: Incubation, Cell Characterization, CyQUANT Assay, Staining, Concentration Assay, Comparison, MANN-WHITNEY

Journal: bioRxiv

Article Title: Urban PM 2.5 at Realistic Environmental Concentrations Impairs Blood–Brain Barrier Integrity and Enhances LOX-1 Expression in Human Brain Endothelial Cells

doi: 10.64898/2026.01.29.702473

Figure Lengend Snippet: Western Blot assessment of adult male HBMEC exposed to vehicle, 5, 15, 75, or 300 μg/m 3 PM 2.5 during normoxia or ischemic-like injury with hypoxia, glucose deprivation and reperfusion (HGD). A . Representative Western Blot image of IL-6 and β-actin band migration. B . Signal quantification of 25kDa IL-6 shows no difference between PM 2.5 exposure or HGD treated group. C . Signal quantification of 17kDa IL-6 shows dose-dependency with higher IL-6 expression from higher PM 2.5 exposure, with significant increase ≥75 μg/m 3 and from HGD treatment compared to vehicle. D . Representative Western Blot image of LOX-1 and β-actin. E . Signal quantification of LOX-1 displays a dose-dependent increase in LOX-1 with exposure to ≥15 μg/m 3 PM 2.5 or HGD. (n=4-7 technical replicates). Data presented as mean +-SD. Statistical significance assessed by Kruskal-Wallis test. *p<0.05, **p<0.01.

Article Snippet: Primary adult male HBMEC were purchased from Innoprot (Spain, Catalog number: P10361, Lot number: 111224CS).

Techniques: Western Blot, Migration, Expressing

Journal: bioRxiv

Article Title: Plasmodium falciparum diacylglycerol acyltransferase maintains phospholipid homeostasis to regulate sexual differentiation, ER stress, and cytoadhesion

doi: 10.1101/2025.10.10.681613

Figure Lengend Snippet: (a) Schematics of cytoadhesion assay under static and flow conditions. (b, c) Dot plots showing the number of erythrocytes infected with Pfdgat :LoxPint:HA parasites that adhered to 100 human brain microvascular endothelial cells (HBMECs) under static (b) and flow (c) conditions after 6 days of rapamycin treatment. Data are shown as mean ± SD from n = 3 independent biological replicates (b) and from n = 2 independent biological replicates from 3 independent experiments (c). P values were calculated using one-way ANOVA followed by Tukey–Kramer test. BFA, brefeldin. (d) Dot plots showing the number of SBP1 puncta on infected erythrocytes with Pfdagt :LoxPint:HA on 6 and 8 days of rapamycin treatment. P values were calculated using one-way ANOVA followed by Tukey–Kramer test. Representative immunofluorescence images are shown above the corresponding plots. Samples were stained for SBP1 with rabbit anti-SBP1 antibody (red) and for nuclei with DAPI (blue). Scale bars = 2 μm. was created with Biorender.com. Source data are provided as a Source Data file.

Article Snippet: For the static cytoadherence assay, immortalized human brain microvascular endothelial cells (HBMEC) (P10361-IM; Innoprot, Dario, Spain) were cultivated on 13-mm coverslips (Matsunami Glass, Osaka, Japan) in 24-well plates containing Endothelial Cell Medium (P60104; Innoprot, hereafter referred to as HBMEC medium) supplemented with 5% fetal bovine serum (175012; Nichirei Biosciences, Tokyo, Japan).

Techniques: Infection, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Evaluation of the In Vitro Blood–Brain Barrier Transport of Ferula persica L. Bioactive Compounds

doi: 10.3390/ijms26168017

Figure Lengend Snippet: ( A ) Cell viability (%) of HBMEC cells after exposure to different concentrations of F. persica aerial part (AerExt) and root (RootExt) extracts for 24 h. ( B ) HBMEC transendothelial electrical resistance (TEER) changes after AerExt (40 µg mL −1 ) and RootExt (20 µg mL −1 ) treatments at different incubation times (2, 4, and 24 h). ( C ) HBMEC sodium fluorescein (Na-F) paracellular permeability after AerExt (40 µg mL −1 ) and RootExt (20 µg mL −1 ) treatments at different incubation times (2, 4, and 24 h). Asterisks (*) denote statistical differences after T-test between control and extract-treated cells at different incubation times, p < 0.05.

Article Snippet: SH-SY5Y and HBMEC cells were obtained from ATCC® (Rockville, MD, USA).

Techniques: Incubation, Permeability, Control