biotinylated maackia amurensis lectin ii  (Vector Laboratories)

Journal: bioRxiv

Article Title: Sialic acids are a barrier to the entry of non-influenza orthomyxoviruses

doi: 10.64898/2026.01.15.699645

Figure Lengend Snippet: A . Correlations between RNA-seq data for sialyltransferases and infectivity of VSV pseudotypes harbouring WBV, SINUV or OZV GPs, as well as PR8 influenza. Gene expression data were quantified as fragments per kilobase per million (FPKM) and infection rate as the percentage of infected cells. Both metrics were scaled between 0 and 100, transformed as log 2 (x+1), and their Pearson correlation was calculated. Genes for which at least one virus showed a significant ( P < 0.01) correlation are shown. The heatmap depicts the average Pearson correlation from two independent infection assays, as shown in the scale bar. The numbers inside the heatmap represent the levels of significance (1: P < 0.01, 2: P < 0.001, 3: P < 0.0001). VSV pseudotyped with its own glycoprotein was included as a control. B . Examples of scatter plots between infectivity and RNA-seq data for the ST6GALNAC2 and ST3GAL5 genes. Pearson correlation coefficients (r) and their statistical significance ( P ) are shown (ns: not significant). The data points for the SK-MEL-28 cell line are marked in red, and those for IGROV-1 cell line are marked in blue. C . SK-MEL-28 and IGROV-1 cells were incubated with biotin-labelled MAL II and SNA lectins to quantify by flow cytometry α2,3- and α2,6-linked SAs, respectively. Lectin-treated cells were incubated with Streptavidin-Dylight488TM (strep-dye) and the signal emitted by the dye was used to quantify SAs. Left: fluorescence data from two independent experiments. Cells treated without lectin and incubated with the strep-dye (strep-dye only) were used as negative control. A t-test between SK-MEL-28 and IGROV-1 values was carried out using log-transformed data (**: P < 0.01; ***: P < 0.001; ns: P > 0.05). Right: fluorescence level histogram from one representative experiment is shown. Y-axis shows the relative cell count (%) of α2,3- or α2,6-linked SAs-positive cells, and X-axis shows the intensity of fluorescence.

Article Snippet: Biotinylated lectins MAL II ( Maackia Amurensis Lectin II , L-1260-2, Vector labs) and SNA ( Sambucus Nigra Lectin , B-1305-2, Vector labs) that specifically bind to α2,3- and α2,6-SAs, respectively, were added at 4°C for 30 min at a concentration of 20 μg/mL in FACS buffer.

Techniques: RNA Sequencing, Infection, Gene Expression, Transformation Assay, Virus, Control, Incubation, Flow Cytometry, Fluorescence, Negative Control, Cell Characterization

Journal: bioRxiv

Article Title: Sialic acids are a barrier to the entry of non-influenza orthomyxoviruses

doi: 10.64898/2026.01.15.699645

Figure Lengend Snippet: Effect of MALII (α2,3-SA specific) and SNA (α2,6-SA specific) lectins on the infectivity of VSV pseudotypes harbouring WBV, SINUV or OZV GPs, as well as PR8 influenza. VSV pseudotyped with its own glycoprotein was included as a control. Viral entry was measured as the percentage of GFP-positive cells at 18-24 hpi. Three independent assays were carried out for each lectin dose. Right: representative images showing the effects of the combined MALII and SNA treatment at a dose of 100 μg/mL each (scale bar, 1000 µm). Statistical tests comparing the infectivity data at 0 versus 100 μg/mL are provided in the text.

Article Snippet: Biotinylated lectins MAL II ( Maackia Amurensis Lectin II , L-1260-2, Vector labs) and SNA ( Sambucus Nigra Lectin , B-1305-2, Vector labs) that specifically bind to α2,3- and α2,6-SAs, respectively, were added at 4°C for 30 min at a concentration of 20 μg/mL in FACS buffer.

Techniques: Infection, Control

Journal: bioRxiv

Article Title: Bovine-derived influenza A virus (H5N1) shows efficient replication in well-differentiated human nasal epithelial cells without requiring genetic adaptation

doi: 10.64898/2026.01.16.699876

Figure Lengend Snippet: a - d HNE ( a ) and HBE ( c ) cells were grown at air-liquid interface at 33 °C and 37 °C, respectively. The cells were labeled using either an antibody directed against the tight junction protein ZO-1 (TJ), lectins MAL-I/MAL-II for staining of α2,3-linked sialic acid and lectin SNA for detection of α2,6-sialic acids. Nuclei were stained with DAPI. Scale bar: 100 µm. Relative number of cells labeled with MAL-I/MAL-II and SNA lectins in HNE ( b ) and HBE ( d ) were determined using ImageJ. Four to six images per sample were analyzed, each containing >100 cells. Mean values and SD are shown. Statistical analysis was performed with the unpaired, two-tailed Student’s t -test.

Article Snippet: For detection of glycoconjugates containing α2,3-linked or α2,6-linked sialic acids, formalin-fixed, non-permeabilized cells were incubated overnight at 4 °C with biotinylated Sambucus nigra agglutinin (SNA, 1:200; Vector Labs, Newark, CA, USA, cat. no. B-1305-2) or combined biotinylated Maackia amurensis lectins I and II (Mal-I, Mal-II, 1:200 each; Vector Labs, cat nos.

Techniques: Labeling, Staining, Two Tailed Test