Journal: bioRxiv
Article Title: Studying macromolecular composition in cell-cell interfaces using 3D membrane reconstitution systems
doi: 10.1101/2025.11.01.686034
Figure Lengend Snippet: Schematic representation of the lectins used (AAL, SNA, WGA, MAL-II) to target different glycosylation patterns in the T cell glycocalyx and their distribution at the T cell GUV contact. B-E| Representative confocal image of contact formation between GUVs decorated with CD2 with Jurkat T cells labelled with WGA (B), AAL (C), SNA (D) and MAL-II (E) (left). CD2-AF488 intensity and WGA-AF647, AAL-AF647, SNA-AF647 or MAL-II-AF647 intensities at the contact and outside were quantified and normalized (middle). Linearized fluorescence intensity profiles of the GUV and cell stained with the respective lectin in the confocal image with the contact highlighted in grey (right). Superplots show individual contacts as small symbols. Large symbols depict the mean of the individual biological replicates. Symbol corresponds to the individual biological replicate (n=3). Standard error of the mean is shown. Scale bar corresponds to 10 μm.
Article Snippet: We also used the proteins: CD43/ human leukosialin (His-Tag, CD3-H52H9, ACROBiosystems), MUC1-Alexa FluorTM 488 (kindly provided by Carolyn Shurer), MHC class I H-2D b presenting the LCMV-derived gp33 peptide (KAVYNFATM) ( ; ), MHC class I (HLA-A*02:01) presenting the tumour-associated antigen NY-ESO1-9V peptide (SLLMWITQV; from now on 9V) , MHC class I (HLA-A*02:01) presenting the NY-ESO1-3P9V peptide variant (SLPMWITQV; from now on 3P9V), P14 TCR , 1G4 TCR , Wheat Germ Agglutinin (WGA) Alexa FluorTM 647 Conjugate (ThermoFisherScientific), Aleuria Aurantia Lectin (AAL) (L-1390-2, Vector Labs), Sambucus Nigra-I-Agglutinin (SNA) (L-1300-5, Vector Labs), Maackia Amurensis-II Lectin (MAL-II) (L-1260-2, Vector Labs).
Techniques: Glycoproteomics, Fluorescence, Staining
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