draq5  (Thermo Fisher)

Journal: Brain Structure & Function

Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system

doi: 10.1007/s00429-026-03088-7

Figure Lengend Snippet: Immunoreactivity of blood vessels and RF to goat anti-PECAM-1 antibody in rat spinal cord. PECAM-1 + (magenta, goat polyclonal anti-PECAM-1 antibody) parenchymal blood vessels (white arrows) and the elongated structure corresponding to RF (white arrowheads) present in the CC of spinal cord ( a , Z-stack); the yellow dashed line square in ( a ) corresponds to the CC lining region; PECAM-1 + RF (magenta, white arrowheads) is viewed in the single optical section and in orthogonal projections of the Z-stack (yellow crossed lines) ( b ); PECAM-1 + parenchymal blood vessels (magenta, white arrows) and RF (white arrowheads) in the coronal section of spinal cord at P8 ( c ) and M18 ( d ); co-localization of SCO-spondin (cyan, rabbit polyclonal anti-SSPO antibody) and PECAM-1 (magenta) in RF (white arrowhead) magnified in white dashed line rectangles in coronal section of spinal cord ( e ); detail of SSPO + (cyan) and PECAM-1 + (magenta) RF (white arrowhead) in the single optical section and in orthogonal projections of the Z-stack (yellow crossed lines), magnified in white dashed line squares ( f ); co-localization of SCO-spondin (cyan) and PECAM-1 (magenta) in RF (white arrowhead) in the longitudinal section of spinal cord, white dashed lines border the CC lumen ( g , Z-stack); nuclei of cells (gray, DRAQ5) in ( a–g ); ultrathin section of spinal cord shows ependymal lining and the CC lumen with RF (white arrowhead) depicted in the yellow dashed line square ( h ) magnified in ( i ) with gold nanoparticles (magenta arrowheads) corresponding to goat anti-PECAM-1 antibody binding to RF; representative images of lumbar spinal cord of rats at postnatal day 8 (P8); 90 days (P90), and 18 months (M18); scale bar: a = 50 μm; b, e, g = 10 μm; c, d = 20 μm; f = 5 μm, h = 2 μm; i = 100 nm

Article Snippet: To visualize cell nuclei, samples were counterstained with DRAQ5 ® (1:500, Cell Signaling Technology, Leiden, Netherlands, #4084) for 10 min, washed in PBS, transferred to glass slides and cover-slipped using ProLong Gold Antifade Reagent with DAPI (Cell Signaling Technology, #8961).

Techniques: Binding Assay

Journal: Brain Structure & Function

Article Title: Anti-PECAM-1 antibodies: another tool for visualization of Reissner´s fiber and the subcommissural organ in rat central nervous system

doi: 10.1007/s00429-026-03088-7

Figure Lengend Snippet: The apical surface of SCO cells and nearby vasculature is immunoreactive to anti-PECAM-1 antibody. Schematic representation of a sagittal section of the brain with the SCO in the third ventricle (3 V) ( a ); a panoramic view showing ependymal lining in the 3 V with a thread-like structure corresponding to RF (*) lying on the surface of the ependymal cells ( b ); the yellow line rectangle in ( b ) represents the region with extracellular material on the surface of the SCO which corresponds to the initial part of RF (pre-RF, magenta pseudo-color) ( c ); the yellow dashed line rectangle in ( b ) represents detail of RF (*) in 3 V, where RF is covered by cilia of multiciliated ependymal cells (yellow arrows) ( d ); RF (*) in the CC of the spinal cord in contact with several cilia of ependymal cells (yellow arrows) ( e ); PAX6 + nuclei (cyan color, rabbit monoclonal anti-PAX6 antibody) of SCO cells, PC corresponds to posterior commissure ( f ); goat anti-PECAM-1 antibodies (magenta) bind to the apical surface of SCO cells and nearby vasculature (white arrows) in neonatal (P8) ( g ), young adult (P90) ( h ), and senescent (M18) rats ( i ); nuclei of cells (gray, DRAQ5) in ( f–i ). Representative images from various age groups (P8, P29, P90, and M18); scale bar: b = 100 μm; c, d = 10 μm; e = 1 μm; f = 20 μm; g = 50 μm, h = 20 μm, i = 50 μm

Article Snippet: To visualize cell nuclei, samples were counterstained with DRAQ5 ® (1:500, Cell Signaling Technology, Leiden, Netherlands, #4084) for 10 min, washed in PBS, transferred to glass slides and cover-slipped using ProLong Gold Antifade Reagent with DAPI (Cell Signaling Technology, #8961).

Techniques:

Journal: bioRxiv

Article Title: Cholesterol remodels the endoplasmic reticulum to control myofibroblastic CAF function

doi: 10.64898/2026.02.16.706237

Figure Lengend Snippet: (A) Measurement of the relative incorporation of [U- 14 C]glucose into protein, non-polar, RNA, DNA and polar fractions following 24 hours of labelling, n=3. (B) Relative incorporation of [1- 14 C]acetate into the lipid fraction of NFs and CAFs, n=3. (C) Gene Set Enrichment Analysis (GSEA) plot derived from RNA-Seq analysis of CAFs versus NFs, showing enrichment of a “lipid metabolic process” signature (GO:0006629). (D) Heatmap of cholesterol biosynthesis gene expression in NFs and CAFs, as determined by RNA-Seq analysis, n=3. (E) Quantification of relative total cholesterol levels in NFs and CAFs, n=3. (F) UMAP visualisation of 19,601 patient-derived breast tumour stromal cells analysed by single cell RNA-Seq (scRNA-Seq) from the Human Breast Cancer Atlas . Stromal clusters corresponding to immunomodulatory CAFs (iCAFs), myCAFs, perivascular (PVL) and endothelial cell subsets are indicated by colour (top). Feature plots illustrating expression of SQLE (bottom left) and SC5D (bottom right) in breast tumour stromal cell clusters. Gene expression is represented by log-normalised expression values. The myCAF cluster is delineated by a dashed line. (G) Representative filipin staining (green) of free cholesterol in NFs and CAFs. Nuclei are stained with Draq5 (blue). Dashed boxes indicate magnified regions shown to the right. White scale bar represents 10 µm, yellow scale bar represents 5 µm. (H) Quantification of relative cholesterol levels in the ER-enriched fraction isolated from NFs and CAFs, n=3.

Article Snippet: Filipin was removed, and cells were washed and imaged in PBS containing 5 μM Draq5 staining solution (Miltenyi Biotec) using a Zeiss Elyra PS.1 confocal microscope (Zeiss) equipped with a 63X/1.4 oil DIC objective.

Techniques: Derivative Assay, RNA Sequencing, Gene Expression, Single Cell, Expressing, Staining, Isolation