Journal: bioRxiv

Article Title: The impact of Staphylococcus aureus cell wall glycosylation on langerin recognition and Langerhans cell activation

doi: 10.1101/2020.11.06.371559

Figure Lengend Snippet: A) Binding of FITC-labeled beads, coated with in vitro glycosylated RboP dodecamers, to muLCs at bead-to-cell ratios of 1, 5, and 10. Bead adherence is displayed as % of FITC+ cells. B) Binding of FITC-labeled beads coated with TarS- or TarP-modified RboP dodecamers to muLCs at a bead-to-cell ratio of 10 in the absence (similar to A) or presence of mannan (20 μg/ml) or anti-langerin blocking antibody (20 μg/ml). C) Binding of FITC-labeled RN4220 Δ tarMS complemented with plasmid-expressed tarS, tarP or tarM to muLCs at a bacteria-to-cell ratio of 1. Bacterial binding is represented by % of FITC+ cells D) Surface expression of activation marker CD86 and maturation marker CD83 by muLCs after 24h stimulation with γ-irradiated RN4220 Δ tarMS complemented with plasmid-expressed tarS, tarP or tarM ,at bacteria-to-cell ratios of 1, 10, and 50. E) Concentration of IL-8 and TNFα in the supernatant of muLCs described in D. The data for all panels represent mean + SEM of biological triplicates. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Cells were washed with PBS 0,1% BSA, stained with CD83 (clone HB15e) and CD86 (clone IT2.2, Sony Biotechnology), fixed and analyzed by flow cytometry.

Techniques: Binding Assay, Labeling, In Vitro, Modification, Blocking Assay, Plasmid Preparation, Bacteria, Expressing, Activation Assay, Marker, Irradiation, Concentration Assay