bend 3 mouse brain endothelial cells  (ATCC)

Journal: Nature Communications

Article Title: Environmental enrichment and physical exercise prevent stress-induced social avoidance and blood-brain barrier alterations via Fgf2

doi: 10.1038/s41467-025-68058-9

Figure Lengend Snippet: A , Experimental timeline for inflammatory insult with TNF-α and FGF2 co-treatment. Once confluent (4 days), HBEC-5i or bEnd.3 were pretreated with FGF2 or vehicle for one hour and then stimulated with TNF-α or vehicle for up to 7 days. FGF2 alters mouse ( B ) and human ( C ) endothelial transcription in response to acute TNF-α stimulation. FGF2 promotes faster restoration of TNF-α-induced Cldn5 loss in mouse ( D ) and human ( E ) endothelial cells. Chronic stimulation with TNF-α leads to a reduction in endothelial monolayer integrity measured by trans-endothelial electrical resistance (TEER) in mouse ( F ) and human ( G ) endothelial cells. FGF2 co-treatment preserves normal TEER despite TNF-α. H 7 days of TNF-α treatment promotes spikes and discontinuities in Cldn5 tight junction strands in bEnd.3, which is reversed by FGF2 treatment (scalebar = 20 μm). Data represent mean ± s.e.m., and each experiment was replicated at least twice on independent samples. Group comparisons were evaluated with two-way ANOVA followed by Bonferroni’s post hoc tests; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The human brain microvascular endothelial cell line HBEC-5i (ATCC CRL-3245, male donor according to https://www.cellosaurus.org/CVCL_4D10 ) and the mouse brain endothelial cell line bEnd.3 (ATCC CRL-2299) were subcultured and stored in banks at −150 ° C. Cells were thawed as needed and cultured in DMEM/F12 supplemented with 10% fetal bovine serum, 25 ug/mL gentamicin (Gibco), and 1X endothelial cell growth supplement (ScienCell).

Techniques:

Journal: bioRxiv

Article Title: Brain endothelial PRMT5-ANGPTL4 axis regulates cerebellar inhibitory synaptogenesis and motor coordination

doi: 10.1101/2025.11.18.688985

Figure Lengend Snippet: (A) UMAP visualization of scRNA-seq data from cerebellar ECs in P20 Prmt5 fl/+ and Prmt5 fl/fl mice, with colors corresponding to the identified clusters. (B) Dot plots showing the expression of specific marker genes in each cluster. (C) Count of significant DEGs in each cluster, with upregulation indicated in orange and downregulation in blue (average |log2FC| > 0.5, adjusted p-value < 0.05). (D) Representative gene ontology (cellular component) analysis of endothelial clusters. Dot size represents the overlap proportion between DEGs and reference gene sets; color intensity reflects -log10 (q-value). (E) Volcano plot showing DEGs encoding ECM components across 6 endothelial clusters. The top 5 genes with the greatest expression differences are labeled. (F) Venn diagram showing intersections among the top 5 genes across 6 endothelial clusters, with overlaps representing shared gene counts. (G) qPCR analysis of Angptl4 mRNA levels in brain ECs derived from P20 Prmt5 fl/+ and Prmt5 fl/fl mice. **P < 0.01 (mean ± SEM, n = 3 per group). (H) Western blot analysis of ANGPTL4 protein expression in brain ECs derived from P20 Prmt5 fl/+ and Prmt5 fl/fl mice. (I) ELISA analysis of secreted ANGPTL4 protein levels. **P < 0.01 (mean ± SEM, n = 3 per group). (J) ChIP-qPCR analysis of PRMT5 occupancy on the Angptl4 promoter region in bEnd.3 cells. ****P < 0.0001 (mean ± SEM, n = 3 per group). (K) qPCR analysis of Prmt5 and Angptl4 mRNA levels in bEnd.3 cells transfected with siNC or siPrmt5 . ***P < 0.001 (mean ± SEM, n = 3 per group). (L) Western blot analysis of PRMT5 and ANGPTL4 protein levels in bEnd.3 cells transfected with siNC or siPrmt5 . GAPDH served as the loading control. (M-O) ChIP-qPCR analysis of the enrichment of H3R8me2s (M), H4R3me2s (N) and H3K9ac (O) at the Angptl4 promoter region in bEnd.3 cells transfected with siNC or siPrmt5 . **P < 0.01, ****P < 0.0001 (mean ± SEM, n = 3 per group).

Article Snippet: Mouse embryonic fibroblasts (NIH-3T3 cells, ATCC, CRL-1658) and mouse brain endothelial cells (bEnd.3 cells, ATCC, CRL-2299) were cultured at 37 °C in a humidified atmosphere containing 5% CO2.

Techniques: Expressing, Marker, Labeling, Derivative Assay, Western Blot, Enzyme-linked Immunosorbent Assay, ChIP-qPCR, Transfection, Control