maackia amurensis lectin ii  (Vector Laboratories)

Journal: Journal of Biomedical Research

Article Title: Generation of ST6GAL1/ST3GAL4 double-knockout Landrace pigs with reduced respiratory sialylated glycans and decreased influenza A virus susceptibility of derived fibroblasts in vitro

doi: 10.7555/JBR.39.20250300

Figure Lengend Snippet: Restoration of influenza virus susceptibility in ST6GAL1 −/− / ST3GAL4 −/− Landrace pig cells through complementation. A: Quantitative reverse transcription-PCR (RT-qPCR) analysis of ST6GAL1 and ST3GAL4 gene expression levels in wild-type (WT), knockout (KO), and complemented (KO + ST) cells. B: Immunofluorescent detection of α2,6-linked and α2,3-linked sialylated glycans in wild-type (WT), knockout (KO), and complemented (KO + ST) Landrace pig cells. Red indicates SNA (α2,6) or MAL Ⅱ (α2,3) staining, and blue indicates nuclei. The KO group showed nearly undetectable signals for both SNA and MAL Ⅱ. In contrast, the WT and KO + ST groups exhibited strong fluorescence for both lectins, indicating successful restoration of both α2,6- and α2,3-linked sialylated glycans in complemented cells. Scale bars, 100 μm. C: Cytopathic effects induced by H1N1 influenza virus infection at 0, 24, and 48 h post-infection (HPI). Scale bars, 500 μm. D: Immunofluorescent detection of IAV nucleoprotein (NP) in cells post-infection with H1N1 influenza virus at 48 HPI. NP-Alexa Fluor 488, green; DAPI, blue. Scale bars, 100 μm. E: RT-qPCR analysis of influenza viral M gene expression levels. F: Plaque assay measurement of infectious IAV (H1N1) virus titers (lg PFU/mL). Data are presented as mean ± standard deviation (A, E, and F); n = 3 independent cell culture experiments conducted on separate days with identical cell batches. Statistical significance was determined by one-way ANOVA followed by Tukey's multiple comparisons test. * P < 0.05, ** P < 0.01, and *** P < 0.001.

Article Snippet: Cells and tissues were then incubated with biotinylated lectins MAL II (1∶200, Cat. #B-1265-1, Vector Laboratories, Burlingame, CA, USA) or SNA (1∶200, Cat. #B-1305-2, Vector Laboratories) for 1 h, followed by three washes with PBS containing 0.05% Tween-20 (PBST).

Techniques: Virus, Reverse Transcription, Quantitative RT-PCR, Analysis, Gene Expression, Knock-Out, Staining, Fluorescence, Infection, Plaque Assay, Standard Deviation, Cell Culture

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