Figure 1. LecB binds to endothelial cells in the skin and obstructs cell emigration.

• A
  Flow cytometry gating strategy to identify the leukocytes emigrating from human skin explants into cell culture medium.
• B
  Human skin explants were cultured in complete medium for 3 days in the absence (nontreated, NT) or presence of LecB or LecA, emigrated cells counted and identified by flow cytometry (panel A).
• C
  As for panel (B) with DH445 and/or LecB added to the culture medium, normalized to the untreated condition.
• D
  Epifluorescence imaging of human skin cross‐sections after incubation of skin explant in culture with LecB‐A488 and then stained for CD31, HLA‐DR and cell nuclei (DAPI). Arrows point to CD31⁺ endothelial cells bound by LecB‐A488.
• E
  Flow cytometry of LecB‐A488 binding to ex vivo isolated CD31⁺ CD146⁺ podoplanin (Pdpn)⁻ blood and CD31⁺ CD146⁻ Pdpn⁺ lymphatic endothelial cells in the absence or presence of different concentrations of the LecB inhibitor DH445.

Data information: data are expressed as mean ± SEM of individual skin donors, that are linked in (C). The data in (C) are normalized to the untreated (NT) condition. ns, not significant, *P < 0.05, ***P < 0.001, as assessed by the Kruskall–Wallis test (B) and Friedman test (C). The scale bars represent 100 μm (D).