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Endothelial cells use dynamic actin to facilitate lymphocyte transendothelial migration and maintain the monolayer barrier.

Mooren OL, Li J, Nawas J, Cooper JA - Mol. Biol. Cell (2014)

Bottom Line: The actin cytoskeleton of the endothelial cell (EC) is known to facilitate transmigration, but the cellular and molecular mechanisms are not well understood.We found that docking structure formation involves the localization and activation of Arp2/3 complex by WAVE2.Finally, we found that ECs in resting endothelial monolayers use lamellipodial protrusions dependent on WAVE2 to form and maintain contacts and junctions between cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Physiology, Washington University, St. Louis, MO 63110.

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WAVE2 promotes VE-cadherin expression and maturation of cell–cell junctions. (A) WAVE2-knockdown monolayers have large gaps between cells, outlined in yellow, and the intensity of VE-cadherin staining is substantially decreased. Scale bar, 50 μm. (B) Decreased total VE-cadherin in WAVE2-knockdown monolayers, by immunoblot. (C) Depletion of WAVE2 leads to a change from mature junctions, collinear with the cell edge, to immature junctions perpendicular to the cell edge, based on staining for VE-cadherin and vinculin. Scale bar, 50 μm.
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Figure 6: WAVE2 promotes VE-cadherin expression and maturation of cell–cell junctions. (A) WAVE2-knockdown monolayers have large gaps between cells, outlined in yellow, and the intensity of VE-cadherin staining is substantially decreased. Scale bar, 50 μm. (B) Decreased total VE-cadherin in WAVE2-knockdown monolayers, by immunoblot. (C) Depletion of WAVE2 leads to a change from mature junctions, collinear with the cell edge, to immature junctions perpendicular to the cell edge, based on staining for VE-cadherin and vinculin. Scale bar, 50 μm.

Mentions: To investigate the molecular basis for the loss of monolayer integrity upon depletion of WAVE2, we examined the molecular composition of cell contacts and cell–cell junctions using immunofluorescence. In control monolayers, VE-cadherin was predominantly organized in a pattern typical for mature junctions—continuous linear structures coinciding with cell–cell contacts (Figure 6A). In contrast, WAVE-depleted monolayers showed very few VE-cadherin–positive cell junctions (Figure 6A). The number of gaps in the junctions increased from 3.0 ± 1.0/fov (mean ± SD, N = 6 fov) in control monolayers to 11.2 ± 2.3/fov in WAVE2-knockdown monolayers. This difference was statistically significant, with p < 0.001.


Endothelial cells use dynamic actin to facilitate lymphocyte transendothelial migration and maintain the monolayer barrier.

Mooren OL, Li J, Nawas J, Cooper JA - Mol. Biol. Cell (2014)

WAVE2 promotes VE-cadherin expression and maturation of cell–cell junctions. (A) WAVE2-knockdown monolayers have large gaps between cells, outlined in yellow, and the intensity of VE-cadherin staining is substantially decreased. Scale bar, 50 μm. (B) Decreased total VE-cadherin in WAVE2-knockdown monolayers, by immunoblot. (C) Depletion of WAVE2 leads to a change from mature junctions, collinear with the cell edge, to immature junctions perpendicular to the cell edge, based on staining for VE-cadherin and vinculin. Scale bar, 50 μm.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4263454&req=5

Figure 6: WAVE2 promotes VE-cadherin expression and maturation of cell–cell junctions. (A) WAVE2-knockdown monolayers have large gaps between cells, outlined in yellow, and the intensity of VE-cadherin staining is substantially decreased. Scale bar, 50 μm. (B) Decreased total VE-cadherin in WAVE2-knockdown monolayers, by immunoblot. (C) Depletion of WAVE2 leads to a change from mature junctions, collinear with the cell edge, to immature junctions perpendicular to the cell edge, based on staining for VE-cadherin and vinculin. Scale bar, 50 μm.
Mentions: To investigate the molecular basis for the loss of monolayer integrity upon depletion of WAVE2, we examined the molecular composition of cell contacts and cell–cell junctions using immunofluorescence. In control monolayers, VE-cadherin was predominantly organized in a pattern typical for mature junctions—continuous linear structures coinciding with cell–cell contacts (Figure 6A). In contrast, WAVE-depleted monolayers showed very few VE-cadherin–positive cell junctions (Figure 6A). The number of gaps in the junctions increased from 3.0 ± 1.0/fov (mean ± SD, N = 6 fov) in control monolayers to 11.2 ± 2.3/fov in WAVE2-knockdown monolayers. This difference was statistically significant, with p < 0.001.

Bottom Line: The actin cytoskeleton of the endothelial cell (EC) is known to facilitate transmigration, but the cellular and molecular mechanisms are not well understood.We found that docking structure formation involves the localization and activation of Arp2/3 complex by WAVE2.Finally, we found that ECs in resting endothelial monolayers use lamellipodial protrusions dependent on WAVE2 to form and maintain contacts and junctions between cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology and Physiology, Washington University, St. Louis, MO 63110.

Show MeSH
Related in: MedlinePlus