Figure 4. PECAM‐1 mediated leukocyte diapedesis is in vivo and in vitro dependent on VE‐cadherin‐Y731.

• A
  Transmigration of mouse neutrophils toward CXCL‐1 through TNF‐α‐stimulated bEnd.5 cells transfected with control or SHP2‐specific siRNA and pre‐treated with anti‐endomucin (7C7.1) or anti‐PECAM‐1 (Mec13.3) antibodies for 30 min at 37°C before addition of neutrophils. The transmigration rate is presented relative to that of control cells, set as 100%.
• B
  Transmigration of mouse neutrophils toward CXCL‐1 through TNF‐α‐stimulated WT or Y731F primary mouse endothelial cells pre‐treated with isotype control or anti‐PECAM‐1 (2H8) antibodies for 30 min at 37°C prior to addition of neutrophils. The transmigration rate is presented relative to that of control cells, set as 100%.
• C–E
  Extravasated leukocytes (C), adherent leukocytes (D), and rolling flux fraction of leukocytes (E) in cremaster tissue from WT or Y731F VE‐cadherin mice stimulated intrascrotally with IL‐1β and treated i.v. with isotype control or anti‐PECAM‐1 (2H8) antibodies for 4 h before intravital microscopy.

Data information: Graphs represent data from three (A, B) independent experiments in triplicate for each group (mean ± SEM). Data in (C–E) are of 38 vessels from four mice, 39 vessels from four mice, 31 vessels from three mice and 43 vessels from four mice analyzed in WT + IgG, WT + 2H8, Y731F + IgG, and Y731F + 2H8 treatment groups, respectively (mean ± SEM). Statistical significance was tested with one‐way ANOVA, *P < 0.05, **P < 0.01, ****P < 0.0001, n.s., not significant.