Fig. 9 | Laboratory Investigation

Fig. 9

From: Preferential expression of sialyl 6′-sulfo N-acetyllactosamine-capped O-glycans on high endothelial venules in human peripheral lymph nodes

Fig. 9

Preferential decoration of CD34 proteins with 297-11A-positive sulfated glycans as compared to mucosal addressin cell adhesion molecule 1 (MAdCAM-1) proteins. a MAdCAM-1 is expressed preferentially on high endothelial venules (HEVs) in Peyer’s patches (PPs) and to a lesser extent in mesenteric lymph nodes (MLNs) but is not expressed in peripheral LNs (PLNs). For this analysis, the same tissue sections used in Fig. 6 were subjected to immunostaining for CD34 (upper panels) or MAdCAM-1 (lower panels). Signals were visualized with 3,3′-diaminobenzidine (brown), and tissues were counterstained with hematoxylin. Bar = 40 μm. b Quantitative immunohistochemical analysis of a total of 216 human LNs showing the percentage of MAdCAM-1-positive HEVs in PLNs (n = 74) and MLNs (n = 142). Data are presented as means plus SEM. ***P < 0.001. c Double immunofluorescence of MLNs for 297-11A (green) and indicated scaffold proteins (red), namely CD34 (upper panels) or MAdCAM-1 (lower panels). Yellow signals in merged panels indicate antigen co-localization. Bar = 20 μm. d Western blot analysis of recombinant CD34•IgG-FLAG and MAdCAM-1•IgG-FLAG proteins decorated with 297-11A-positive sulfated glycans. Membrane was immunoblotted (IB’d) with anti-FLAG M2 (left panel) or 297-11A (right panel)

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