Extended Data Fig. 8: Extended characteristics of GlialCAMAA370-389 and immunofluorescence stainings with MS39p2w174.
From: Clonally expanded B cells in multiple sclerosis bind EBV EBNA1 and GlialCAM
a, Phage display PhiP-Seq data, showing alignment of Pro/Arg-rich region and adjacent residues of all phage display peptides enriched above 100 / 105 reads. b, Immunofluorescence of mouse brain slices stained with (i) control antibody, and (ii-iv) MS39p2w174 (green) and DAPI (blue). (i,ii) full brain, scale bars: 2000 µm, (iii) magnification of hippocampus with prominent MS39p2w174 staining, scale bar: 400 µm, and (iv) olfactory bulb with prominent MS39p2w174 staining in the olfactory nerve (oln), glomerular (gl), and external plexiform layers (epl), but not the mitral (ml), internal plexiform (ipl), or granule cell (gcl) layers, scale bar: 100 µm. c, Immunofluorescence of primary rat oligodendrocytes with isotype control antibody (top panel) and MS39p2w174 (bottom panel). d, K562 cells in culture, wildtype (left) and transduced with full-length GlialCAM (right). e, Immunofluorescence with MS39p2w174 on WT K562 cells (top) and GlialCAM-tg K562 cells (center and bottom). White arrow: single K562 cell, orange arrow: high intensity MS39p2w174 staining on the cell border between transgenic K562 cells in bulks. c, e, Scale bars: 40 µm. b–e, representative micrographs of at least two experiments. f, Overview of phosphorylated residues in GlialCAM, identified by mass spectrometry (phosphoSite.org). The two phosphorylated serine residues of interest are indicated with arrows. g, ELISA, measuring binding of MS39p2w174 to native and citrullinated GlialCAMAA370-389 peptides, means of n = 2 independent experiments, each carried out in triplicates. Wildtype, WT; extracellular domain, ECD; intracellular domain, ICD; phosphorylated serine, pSer; citrulline residue, Cit.
