Journal: Frontiers in Pharmacology

Article Title: Mycobacterium tuberculosis infection drives osteoclast overactivation via α2,3-Sialylation to promote pathological bone destruction

doi: 10.3389/fphar.2026.1738896

Figure Lengend Snippet: α2,3-sialylation is required for BCG-induced osteoclast differentiation and activity (A) Comparison of differentially expressed genes (DEGs) between BCG-infected osteoclasts (RB) and uninfected controls (R). (n = 3 biological replicates per group, differential expression was defined as |FoldChange| > 2 and padj <0.05). (B) Volcano plot of DEGs highlighting genes related to sialic acid biosynthesis. (C) KEGG pathway enrichment analysis of upregulated DEGs in RB cells. (D,E) Immunofluorescence staining of α2,3-SA in mouse calvarial sections using MAL II lectin, with quantification of α2,3-SA fluorescence intensity (n = 5). Intensity density was normalized to the PBS group mean. (F) In vitro osteoclasts subjected to MAL II lectin staining and TRAP staining, with or without sialidase treatment to enzymatically remove α2,3-SA. (G) Quantification of α2,3-SA fluorescence intensity in cultured osteoclasts (n = 5). Intensity density was measured in cellular ROIs after background subtraction and normalized to the RANKL group mean. (H) Quantification of TRAP + multinucleated cells (≥3 nuclei) per field (randomly selected fields, fixed magnification) in vitro (n = 5). (I) mRNA expression levels of osteoclast differentiation markers ( Fos, Mmp9, Nfatc1, and Ocstamp ) in osteoclasts (n = 3). Data are presented as mean ± SD. Statistical significance was determined by two-tailed unpaired Student’s t-test for two-group comparisons (E) and one-way ANOVA followed by Tukey’s post hoc test for three-group comparisons (G–I) .

Article Snippet: Biotin MAL-II , Vector laboratories , Cat# B-1265-1.

Techniques: Activity Assay, Comparison, Infection, Quantitative Proteomics, Immunofluorescence, Staining, Fluorescence, In Vitro, Cell Culture, Expressing, Two Tailed Test

Journal: Scientific Reports

Article Title: In Silico Prediction and Experimental Confirmation of HA Residues Conferring Enhanced Human Receptor Specificity of H5N1 Influenza A Viruses

doi: 10.1038/srep11434

Figure Lengend Snippet: a,b) The binding of varying amounts of biotinylated lectins MAAII (specific for avian α2,3 receptors) and SNA/EBL (specific for human α2,6 receptors) to QT6 and MDCK-SIAT1 cells as well as A549 control cells was determined by FACS analysis using streptavidine-PE. The differences of binding of varying amounts of biotinylated lectins to different cell lines were statistically compared to QT6 for MAAII-lectin binding or A549 for SNA/EBL-lectin binding using two-way ANOVA, followed by Bonferroni post hoc test. Significant differences between data sets are indicated by asterisks ( * ), whilst (ns) indicates non-significant differences. c ) Formaldehyde-fixed monolayers of the indicated cell types were incubated with fluorescein-labelled SNA lectin to detect α2,6-linked sialic acids and biotinylated MAAII lectin and streptavidin-Cy3 to detect α2,3-linked sialic acids. The cellular membrane was permeabilized, the nuclei were stained with DAPI and analyzed using confocal microscopy.

Article Snippet: Cells were incubated for 1 h at RT with fluorescein-labelled Sambucus nigra agglutinin (SNA, Biozol, Germany, 1:100 dilution) to detect α2,6-SA or with biotinylated Maackia amurensis lectin II (MAAII, Biozol, Germany, 1:100 dilution) to detect α2,3-SA.

Techniques: Binding Assay, Control, Incubation, Membrane, Staining, Confocal Microscopy