Figure 3 | Scientific Reports

Figure 3

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

Figure 3

Determination of α2,6- and α2,3 receptors on MDCK-SIAT1, A549 and QT6 cells with specific lectins.

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.

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