Journal: bioRxiv

Article Title: Single-Cell and Spatial Methods for Multimodal Functional Glycan Profiling in Tissues

doi: 10.64898/2026.02.23.707224

Figure Lengend Snippet: (A) Dot plot showing the expression of different marker genes used for cell type annotation across different cell types. (B-C) Violin plots depicting log-normalized expression of lectin-binding ligands for Siglec-7L, Siglec-9L, Siglec-15L, DC-SIGNL, and Galectin-8L across cell types and perturbations. (D) Scatter density plot showing different populations of TCR stimulated CD8+ T cells and NK cells binned by expression levels of Siglec-9L and Siglec-15L. (E) Heatmap of key functional CD8+ T cell and NK cell gene expression across TCR stimulated CD8+ T cells and NK cells grouped by expression levels of Siglec-9L and Siglec-15L. (F) Heatmap showing Pearson correlation of functional CD8+ T cell activation genes with Siglec-7L, Siglec9L and Siglec-15L (top) and their expression across CD8+ T cells in different perturbations (bottom). (G) Expression and clustering of different lectin-ligands across different cell types and conditions. (H) Expression and clustering of O-glycosylation related genes across different cell types and conditions.

Article Snippet: We defined the follicular center as the region located 80-150 μm inward from the Galectin-8L High mantle zone rim, the mantle zone as spanning 0-80 μm, and the marginal zone as extending outward of the rim.

Techniques: Expressing, Marker, Binding Assay, Functional Assay, Gene Expression, Activation Assay, Glycoproteomics

Journal: bioRxiv

Article Title: Single-Cell and Spatial Methods for Multimodal Functional Glycan Profiling in Tissues

doi: 10.64898/2026.02.23.707224

Figure Lengend Snippet: (A) Dot plot showing the expression of different marker genes used for cell type annotation across different cell types. (B) Violin plots depicting log-normalized expression of lectin-binding ligands for DC-SIGNL, Galectin-8L, MBL-L and Siglec-15L across cell types and perturbations. (C) Expression and clustering of O-glycosylation related genes across different cell types and conditions. (D) Heatmap of key functional CD4+ T gene expression across perturbations. (E) GSEA analyses across CD4+ T between LPS stimulation with anti-Siglec7 and anti-Siglec9 blocking antibodies compared to LPS treatment alone.

Article Snippet: We defined the follicular center as the region located 80-150 μm inward from the Galectin-8L High mantle zone rim, the mantle zone as spanning 0-80 μm, and the marginal zone as extending outward of the rim.

Techniques: Expressing, Marker, Binding Assay, Glycoproteomics, Functional Assay, Gene Expression, Blocking Assay

Journal: bioRxiv

Article Title: Single-Cell and Spatial Methods for Multimodal Functional Glycan Profiling in Tissues

doi: 10.64898/2026.02.23.707224

Figure Lengend Snippet: (A) Schematic workflow of GlycoScope, illustrating the sequential staining of antibody and recombinant lectin panels. (B) Representative multiplexed images showing nuclei (DAPI), Siglec-9L, Siglec-15L, Galectin-8L, T cells (CD3), B cells (CD20), and follicular dendritic cells (CD21) across GlycoScope, Antibody-only, and Lectin-only conditions of tonsil tissues (scale bar: 50µm, 10µm (inset)). All individual images across these three conditions are in . (C) Heatmap showing the 0,1 normalization of mean expression (left) and z-score of the coefficient of variation (right) of all glycan and protein markers obtained from GlycoScope, Antibody-only, and Lectin-only conditions. (D) Schematic overview of the GlycoScope study cohorts. Cohort 1 consists of an FFPE tissue microarray (TMA) containing four tonsil, four follicular hyperplasia (FH), and eight follicular lymphoma (FL) cores, profiled using a 32-plex protein panel and six oligo-conjugated lectins (Siglec-7, Siglec-9, Siglec-15, MBL, DC-SIGN, and Galectin-8). Cohort 2 consists of an FFPE TMA containing 11 FL and 5 FH samples, profiled using a 42-plex protein panel and six oligo-conjugated lectins (Siglec-7, Siglec-9, Siglec-15, DC-SIGN, Galectin-8, and Galectin-9). (E) Representative GlycoScope multiplexed images (top) showing nuclei (DAPI), B/tumor cells (Pax5), macro-phages (CD68), T cells (CD3), endothelial cells (CD31), FDC-enriched region (CD21) and lymphatic (Podoplanin), as well as the corresponding phenotype maps (bottom) of FL, FH and tonsil tissues (scale bar: 100µm, 20µm (inset)). All individual phenotype maps for each tissue sample core are in . (F) Top: Representative GlycoScope multiplexed images of FL and FH showing Siglec-7L (magenta), CD31 (yellow), and PAX5 (cyan). Bottom: Representative GlycoScope multiplexed images of FL and FH showing Siglec-9L (cyan), CD31 (magenta), and PAX5 (white). Scale bar, 100 µm. (G) Representative GlycoScope multiplexed images of FL and FH showing DC-SIGNL (cyan), CD21 (magenta) and CD21 (yellow). scale bar: 100µm. (H) Violin plots showing the expression of DC-SIGNL acquired from GlycoScope in tonsil, FH and FL tissues (Krus-kal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (I) Histogram showing the expression distribution of DC-SIGNL in B cell. (J) Violin plots showing the expression of BCL2 protein in DC-SIGNL High and DC-SIGNL Low B cells (Pairwise Wilcoxon test). (K) Line plot showing the mean expression of DC-SIGNL in follicle B cell at 0-150 µm from the anchored FDC. ***P<0.001; ****P<0.0001

Article Snippet: We defined the follicular center as the region located 80-150 μm inward from the Galectin-8L High mantle zone rim, the mantle zone as spanning 0-80 μm, and the marginal zone as extending outward of the rim.

Techniques: Staining, Recombinant, Expressing, Glycoproteomics, Microarray

Journal: bioRxiv

Article Title: Single-Cell and Spatial Methods for Multimodal Functional Glycan Profiling in Tissues

doi: 10.64898/2026.02.23.707224

Figure Lengend Snippet: (A) Violin plots showing the median expression of Galectin-8L across cell types (Kruskal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (B) Representative GlycoScope multiplexed images with markers for Galectin-8L and B cells (Pax5) shown (scale bar: 20µm). (C) Violin plots showing the mean expression of Galectin-8L in B cells within the follicle across tonsil, FH, and FL tissues (Kruskal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (D) Mean expression of Galectin-8L in BCL6+, BCL2+, and neo-plastic BCL2+BCL6+ follicular B cells across tonsil, FH, and FL tissues. (E) Representative GlycoScope multiplexed images with markers for Galectin-8L (green), BCL6 (pink), and CD3 (cyan) shown (scale bar: 100µm). White dotted lines marked the regions for mantle zone, follicle center and neoplastic follicle. (F) Representative GlycoScope multiplexed images with markers for Galectin-9L (magenta), BCL6 (yellow), and CD3 (cyan) shown (scale bar: 100µm). White dotted lines marked the regions for follicle center and neoplastic follicle. (G) Representative GlycoScope multiplexed images with markers for Galectin-8L (magenta), Galectin-8 (yellow), and CD3 (cyan) shown (scale bar: 100µm). White dotted lines marked the regions for mantle zone, follicle center and neoplastic follicle. (H) Schematic illustration of stepwise extension analysis. Galectin-8L+ B cells at the mantle-marginal zone rim were anchored and expanded inward towards the follicle center (blue) and outward towards marginal zone (red), in 10 µm radial steps. (I) Mean proportions of Treg between 0-140µm towards the follicle center and marginal zone from the anchor rim are shown. The region in grey symbolizes the follicle. Mean proportions of FDC-enriched B cells, CD4+ T, CD8+ T, CD68+ macrophage and CD68+CD163+ macrophages are shown in (Kruskal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (J) Mean expression of Galectin-8L in follicular B cells at each step across tonsil, FH, and FL tissues. (K) Correlation scatter plot of the proportion of PD1+ (left) and Tox1/2+ (right) T cells with Galectin-8L expression in B cells at each step (distance towards follicle center from anchor rim) (Spearman’s correlation; p-value adjusted with Benjamini-Hochberg method). (L) Top: Correlation scatter plot of the proportion of CD3T cells with Galectin-8L expression in B cells at each step (distance towards follicle center from anchor rim). Bottom: Correlation scatter plot of the proportion of Tox1/2+ or PD1+ T cells with BCL2 expression in B cells at each step (distance towards follicle center from anchor rim) (Spearman’s correlation; p-value adjusted with Benjamini-Hochberg method). (M) Schematic illustration of summary findings. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns P>0.05.

Article Snippet: We defined the follicular center as the region located 80-150 μm inward from the Galectin-8L High mantle zone rim, the mantle zone as spanning 0-80 μm, and the marginal zone as extending outward of the rim.

Techniques: Expressing

Journal: bioRxiv

Article Title: Single-Cell and Spatial Methods for Multimodal Functional Glycan Profiling in Tissues

doi: 10.64898/2026.02.23.707224

Figure Lengend Snippet: (A) Additional multiplexed images of tonsil, FH and FL cores showing Galectin-8L (green), BCL6 (pink) and CD3 (cyan) markers (scale bar: 20µm). (B) Mean proportions of FDC-enriched B cells between 0-140µm towards the follicle center and marginal zone from the anchor rim are shown (Kruskal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (C) Mean proportions of CD8+ T (top left), CD4+ T (top right), CD68+ Macrophage (bottom, left) and CD68+ CD163+Macrophage (bottom right), between 0-140µm towards the follicle center and marginal zone from the anchor rim are shown (Kruskal-Wallis rank sum test followed by pairwise Wilcoxon test with Benjamini-Hochberg adjustments). (D) Correlation scatter plot of the proportion of LAG3+ (left) and TIM3+ (right) T cells with Galectin-8L expression in B cells at each step within the follicle (Spearman’s correlation with Benjamini-Hochberg adjustments). *P<0.05; **P<0.01; ****P<0.0001; ns P>0.05.

Article Snippet: We defined the follicular center as the region located 80-150 μm inward from the Galectin-8L High mantle zone rim, the mantle zone as spanning 0-80 μm, and the marginal zone as extending outward of the rim.

Techniques: Expressing