Journal: bioRxiv

Article Title: Microfluidic analysis reveals ROCK2 regulation of endothelial cilia is essential for blood vessel lumen formation and vascular integrity

doi: 10.64898/2026.05.27.728336

Figure Lengend Snippet: (A) Schematic of a primary cilium, depicting its core compartments (basal body, transition zone, axoneme, ciliary tip) and the localization of key ciliary proteins ARL13B and RPGRIP1L. Intraflagellar transport (IFT) complexes (IFT-A and IFT-B) facilitate cargo trafficking along the axoneme. Adapted from Smith et al. (2020). (B, C) Immunofluorescence images confirming primary cilia (ARL13B, red) in human cerebral microvascular endothelial cells (hCMEC/D3) (B) and mouse bEnd.3 cells (C) . Nuclei are counterstained with DAPI (blue). Scale bars, 50 µm. (D) Primary cilia (ARL13B, red) in a monolayer of human umbilical vein endothelial cells (HUVECs). Nuclei are stained with DAPI (blue). Scale bar, 50 µm. (E) Primary cilia (ARL13B, red) on endothelial tubules (white arrowheads) and fibroblasts (yellow arrowheads) in a HUVEC–human dermal fibroblast (HDF) organotypic coculture. L, lumen. Nuclei are stained with DAPI (blue). Scale bar: 15 µm. (F, G) Endothelial primary cilia in E13.5 mouse embryonic brain vessels. Cilia (arrows, ARL13B, red) are visible on endothelial cells lining the lumen of a superficial vessel (F) and on intraparenchymal vessels (G) . Endomucin (green (F), brown (G)) marks endothelial cells; nuclei are stained with DAPI (blue). Scale bars, 50 µm (F) and 10 µm (G) .

Article Snippet: The following cell lines were cultured in a humidified atmosphere at 37°C with 5% CO 2 . hCMEC/D3 (human cerebral microvascular endothelial cells) (ATCC® CRL-3245TM) were cultured in EndoGRO-MV Complete Media (Merck, SCME004) supplemented with 1 ng/ml basic Fibroblast Growth Factor (bFGF; PeproTech, 100-18B) and used between passages 30-35 for ciliogenesis assays. bEnd.3 (mouse brain endothelial cells) (ATCC® CRL-2299TM) were cultured in DMEM (Gibco, 11995065) supplemented with 10% (v/v) heat-inactivated foetal bovine serum (FBS) (Merck, F9665), 100 U/ml penicillin, 100 μg/ml streptomycin, and 2 mM L-glutamine (all from Gibco). hTERT RPE-1 (human retinal pigment epithelial cells) (ATCC® CRL-4000TM) were cultured in DMEM/F12 (Gibco, 11330032) supplemented with 10% FBS, penicillin/streptomycin, and L-glutamine.

Techniques: Immunofluorescence, Staining

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Establishment and characterization of an inflammatory tricellular BBB in vitro model. ( A ) Changes in TEER of hCMEC/D3 cells before and after LPS treatment, with LPS stimulation significantly reducing TEER levels. ( B ) Permeability assay using FITC-dextran, showing an approximately 2.6-fold increase in permeability following LPS stimulation, indicating barrier function impairment. ( C ) Immunofluorescence reveals MAP2⁺ neuronal processes extending along GFAP⁺ astrocytic end-feet, forming synapse-endfoot contacts. ( D ) Top-down view of cellular co-localization in the tricellular co-culture system, displaying neurons, astrocytes, and endothelial cells co-existing within the same field of view. ( E ) Schematic representation of the spatial distribution within the tricellular in vitro BBB model, illustrating the arrangement of different cells in the Transwell system. ( F ) VE-cadherin immunostaining shows changes in endothelial cell junction integrity before and after LPS treatment, with blurred junctional boundaries and reduced expression post-stimulation. Data are presented as mean ± SEM. Statistical significance in panels A and B was determined using a two-tailed unpaired Student’s t-test, **p < 0.01, ***p < 0.001

Article Snippet: In the inflammatory hCMEC/D3 model, cells were pretreated with the AMPK inhibitor Compound C (10 μM, MCE) for 1 h prior to nanoparticle exposure.

Techniques: In Vitro, Permeability, Immunofluorescence, Co-Culture Assay, Immunostaining, Expressing, Two Tailed Test

Journal: Journal of Nanobiotechnology

Article Title: Application of an intercellular adhesion molecule-1-targeted nanoparticles loaded with irisin in postoperative neurocognitive disorder through metabolic axis activation and barrier restoration

doi: 10.1186/s12951-026-04271-y

Figure Lengend Snippet: Enhanced targeted uptake of ICAM-1-NP@Irisin in inflammatory brain endothelial cells. ( A ) Confocal laser scanning micrographs: Representative fluorescence images of ICAM-1-NP@Irisin and NP@Irisin uptake in inflammatory hCMEC/D3 cells at 1 h, 3 h, and 6 h (red indicates NP fluorescence, blue indicates DAPI nuclear staining, scale bar 25 μm); the ICAM-1-NP@Irisin group exhibits stronger fluorescence signals with higher distribution density. ( B ) Quantitative analysis of fluorescence intensity: At all time points, intracellular fluorescence signals in the ICAM-1-NP@Irisin group are significantly higher than those in the NP@Irisin group (1 h: 1.4 ± 0.1 fold; 3 h: 1.8 ± 0.2 fold; 6 h: 2.1 ± 0.2 fold; data are presented as mean ± SEM. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test, ****p < 0.0001).

Article Snippet: In the inflammatory hCMEC/D3 model, cells were pretreated with the AMPK inhibitor Compound C (10 μM, MCE) for 1 h prior to nanoparticle exposure.

Techniques: Fluorescence, Staining