anti ly6a  (R&D Systems)

Journal: eLife

Article Title: A novel 3D visualization method in mice identifies the periportal lamellar complex (PLC) as a key regulator of hepatic ductal and neuronal branching morphogenesis

doi: 10.7554/eLife.108669

Figure Lengend Snippet: ( A ) UMAP projection of single-cell transcriptomes of liver endothelial cells from normal adult mice reveals 10 distinct cellular clusters. Each dot represents one cell. ( B ) Heatmap showing expression profiles of representative genes across the 10 clusters, including hepatic stellate cells, T cells, macrophages, hepatocytes, cholangiocytes, portal vein ECs, periportal LSECs, midzonal LSECs, pericentral LSECs, and central vein ECs. ( C ) Representative cluster-specific markers for five major liver endothelial subpopulations. ( D ) Spatial schematic illustrating the anatomical position of the PLC, located exclusively between the portal vein ECs and periportal LSECs. ( E ) Venn diagram showing overlapping top 20 highly expressed genes between portal vein ECs and periportal LSECs. ( F ) Multiplex immunofluorescence images showing CD31 (yellow), Sca-1 (green), and CD34 (red) expression in distinct hepatic endothelial zones, including portal vein ECs, midzonal LSECs, and central vein ECs. The PLC structure is demarcated by dashed white arrows. Scale bar: 40 μm. ( G ) Classification of CD34 + Sca-1 + double-positive versus double-negative endothelial subpopulations. ( H ) Volcano plot illustrating differentially expressed hematopoietic-associated genes in CD34 + Sca-1 + cells. The x-axis shows log₂ fold-change (log₂FC); the y-axis shows −log₁₀ adjusted p value. Significantly upregulated genes are located in the upper right quadrant. ( I ) Gene Ontology enrichment analysis of genes upregulated in CD34 + Sca-1 + ECs, highlighting functional categories such as stem cell development, differentiation, and stem cell maintenance. ( J ) The schematic illustration depicts the spatial localization of Sca-1, a marker for mesenchymal or hematopoietic stem cells, and CD34 + Sca-1 + CD31 + endothelial cells as the trunk of the PLC. Figure 4—source data 1. Source data for : Re-analysis of single-cell RNA-seq data from .

Article Snippet: Antibody , Anti-LY6A (Sca-1) (rabbit monoclonal) , HUABIO , Cat# ET1703-67; RRID: AB_3070422 , 1:500.

Techniques: Single Cell, Expressing, Multiplex Assay, Immunofluorescence, Functional Assay, Marker, RNA Sequencing

Journal: eLife

Article Title: A novel 3D visualization method in mice identifies the periportal lamellar complex (PLC) as a key regulator of hepatic ductal and neuronal branching morphogenesis

doi: 10.7554/eLife.108669

Figure Lengend Snippet: ( A ) UMAP dimensionality reduction analysis showing five distinct endothelial cell clusters isolated from normal mouse liver after excluding non-endothelial cells. Each dot represents an individual cell. ( B ) Quantification of the number of cells in the five endothelial subpopulations: central vein ECs, pericentral LSECs, periportal LSECs, midzonal LSECs, and portal vein ECs. The results showed that central vein ECs were the most abundant (669 cells), while portal vein ECs were the least (146 cells). This reflects the spatial quantitative differences among liver endothelial subpopulations based on single-cell RNA sequencing clustering. ( C ) UMAP-based heatmaps displaying the expression patterns of CD34 (left) and Sca-1 (right) across different liver endothelial subpopulations. Both markers were predominantly expressed in portal vein ECs, periportal LSECs, and midzonal LSECs regions. The color gradient indicates expression levels, with deeper red representing higher expression. ( D ) Immunofluorescence staining of CD34, Sca-1, CD36, and PDGFRB in mouse liver tissue sections. CD34 and Sca-1 were primarily localized around portal veins (PV) and adjacent vascular regions. CD36 was predominantly expressed in periportal liver sinusoidal endothelial cells, while PDGFRB showed widespread distribution in multiple intrahepatic endothelial cell populations. CV: central vein; PV: portal vein. Scale bar: 200 μm. ( E ) Multiplex fluorescence imaging and 3D reconstruction of portal vein endothelial cells (ECs). From left to right: Sca-1/CD31/CD34/DAPI fluorescence imaging showing the distribution of Sca-1 (green), CD31 (yellow), CD34 (red), and nuclei (DAPI, blue). Imaris 3D reconstruction visualizing the spatial organization of portal vein ECs. Magnified Sca-1/CD31/CD34/DAPI image highlighting marker distribution at the cellular level. Enlarged 3D reconstruction showing the spatial arrangement of Sca-1 + , CD31 + , and CD34 + cells.

Article Snippet: Antibody , Anti-LY6A (Sca-1) (rabbit monoclonal) , HUABIO , Cat# ET1703-67; RRID: AB_3070422 , 1:500.

Techniques: Isolation, Single Cell, RNA Sequencing, Expressing, Immunofluorescence, Staining, Multiplex Assay, Fluorescence, Imaging, Marker

Journal: eLife

Article Title: A novel 3D visualization method in mice identifies the periportal lamellar complex (PLC) as a key regulator of hepatic ductal and neuronal branching morphogenesis

doi: 10.7554/eLife.108669

Figure Lengend Snippet: ( A ) Multiplex immunofluorescence staining showing the distribution of CD31 (yellow), Sca-1 (green), and CK19 (red) in the liver of control mice. Scale bar: 50 μm. ( B ) Higher-magnification view of multiplex immunofluorescence staining showing CD31 (yellow), Sca-1 (green), and CK19 (red) distribution in control mouse livers. Scale bar: 20 μm. ( C ) Quantification of the terminal bile duct area on the surface of portal veins with different diameters in control mice. Data are presented as mean ± standard deviation (Mean ± SD). Number of portal veins: control = 20, CCl 4 –6 weeks=18. ( D ) Quantification of CD34 + Sca-1 + endothelial cell subpopulations in control and fibrotic (CCl 4 -induced) mouse livers. Figure 5—figure supplement 1—source data 1. Quantification of terminal bile duct area on portal vein surfaces with different diameters in control mice.

Article Snippet: Antibody , Anti-LY6A (Sca-1) (rabbit monoclonal) , HUABIO , Cat# ET1703-67; RRID: AB_3070422 , 1:500.

Techniques: Multiplex Assay, Immunofluorescence, Staining, Control, Standard Deviation

Journal: eLife

Article Title: A novel 3D visualization method in mice identifies the periportal lamellar complex (PLC) as a key regulator of hepatic ductal and neuronal branching morphogenesis

doi: 10.7554/eLife.108669

Figure Lengend Snippet: ( A ) Volcano plot of differentially expressed bile duct-related genes in CD34 + Sca-1 + double-positive cells. The x-axis represents log2 fold change (log2FC), and the y-axis represents -log10 adjusted p value (-log10(p-adjust)). Genes significantly upregulated are located in the upper right quadrant. ( B ) Pathway enrichment analysis of upregulated genes in CD34 + Sca-1 + cells showed enrichment in categories such as epithelial morphogenesis and branching morphogenesis of epithelial tubes, represented as −log₁₀(p value). ( C ) Visualization of portal vein labeled with MCNP-Pink and bile duct epithelial cells stained for CK19 (brown) by 3D DAB immunohistochemistry in control and CCl 4 6-week fibrotic mice. Scale bar: 200 μm. ( D ) Quantification of the distance that bile duct termini extend from portal vein surfaces into hepatic parenchyma in control and CCl 4 6-week mice, presented as mean ± SD (control n=20, CCl 4 6 weeks n=18). ( E ) Multiplex immunofluorescence showing expression and spatial distribution of CD31 (yellow), Sca-1 (green), and CK19 (red) in control and fibrotic models (CCl 4 3 weeks and 6 weeks). White arrows in CK19 single-channel magnified images indicate bile duct termini. Scale bar: 50 μm. ( F ) Quantitative measurement of bile duct termini extension distances along PLC structures into hepatic parenchyma in control, early (CCl 4 3 weeks), and late (CCl 4 6 weeks) fibrosis mice. Data represent mean ± SD (n=5 per group). Statistical significance determined by one-way ANOVA with Tukey’s multiple comparisons test; *p<0.05, **p<0.01, ****p<0.0001. ( G ) Volcano plot of differentially expressed bile duct-related genes in CD34 + Sca-1 + cells from fibrotic livers compared to controls. Axes as in ( A ). Significantly upregulated genes are in the upper right quadrant. ( H ) Schematic illustration showing spatial localization of CK19 and CD31 within the PLC structure. Figure 5—source data 1. Quantification of bile duct termini extension from portal vein surfaces into hepatic parenchyma in control and CCl₄ 6-week mice. Figure 5—source data 2. Quantitative measurement of bile duct termini extension along PLC structures into hepatic parenchyma in control and fibrosis mice at early (CCl₄ 3 weeks) and late (CCl₄ 6 weeks) stages.

Article Snippet: Antibody , Anti-LY6A (Sca-1) (rabbit monoclonal) , HUABIO , Cat# ET1703-67; RRID: AB_3070422 , 1:500.

Techniques: Labeling, Staining, Immunohistochemistry, Control, Multiplex Assay, Immunofluorescence, Expressing

Journal: eLife

Article Title: A novel 3D visualization method in mice identifies the periportal lamellar complex (PLC) as a key regulator of hepatic ductal and neuronal branching morphogenesis

doi: 10.7554/eLife.108669

Figure Lengend Snippet: ( A ) Differential expression analysis of nerve-related genes in CD34 + Sca-1 + endothelial cells. The x-axis indicates log2 fold change (log2FC), and the y-axis represents –log10 adjusted p value (–log10(p-adjust)). Significantly upregulated genes are located in the upper right quadrant. ( B ) Gene Ontology (GO) enrichment analysis of upregulated genes in CD34 + Sca-1 + cells, showing enrichment in functional categories such as semaphorin-plexin mediated axon guidance, regulation of neuronal projection regeneration, and modulation of postsynaptic neurotransmitter receptor endocytosis. Enrichment significance is indicated by –log10(p-value). ( C ) Multiplex immunofluorescence staining showing tyrosine hydroxylase (TH, green) labeling sympathetic nerves and CD31 (yellow) labeling portal vein endothelium. Scale bar: 20 μm. ( D ) Distribution of CD31 (yellow) and TH (green) expression in control and CCl 4 -induced liver fibrosis models at week 3 and week 6, visualized by multiplex immunofluorescence. In the green channel images, the white arrows mark the terminal location of TH-positive sympathetic nerve endings. Scale bar: 50 μm. ( E ) Quantification of the distance from sympathetic nerve endings to the hepatic parenchyma along PLC structures in control, early fibrosis (CCl 4 –3 weeks), and advanced fibrosis (CCl 4 –6 weeks) mice. Data are presented as mean ± standard deviation (Mean ± SD), n=5 per group. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparison test; *p<0.05, **p<0.01, ****p<0.0001. ( F ) Differential expression analysis of nerve-related genes between CD34 + Sca-1 + cells from fibrosis and control livers. The x-axis indicates log2 fold change (log2FC), and the y-axis represents –log10 adjusted p value (–log10(p-adjust)). Significantly upregulated genes are located in the upper right quadrant. ( G ) Schematic diagram illustrating the spatial localization of TH-positive sympathetic nerves and CD31-positive endothelial cells within PLC structures. Figure 6—source data 1. Quantification of the distance from sympathetic nerve endings to hepatic parenchyma along PLC structures in control and fibrosis mice (early and advanced stages).

Article Snippet: Antibody , Anti-LY6A (Sca-1) (rabbit monoclonal) , HUABIO , Cat# ET1703-67; RRID: AB_3070422 , 1:500.

Techniques: Quantitative Proteomics, Functional Assay, Multiplex Assay, Immunofluorescence, Staining, Labeling, Expressing, Control, Standard Deviation, Comparison