Skip to main content
. Author manuscript; available in PMC: 2009 Nov 1.
Published in final edited form as: Cell Commun Adhes. 2008 Nov;15(4):333–349. doi: 10.1080/15419060802440377

Figure 4. Relationship between VE-cadherin and N-cadherin protein levels in BPAEC.

Figure 4

(A) SiRNA sequences targeting either luciferase or N-cadherin were delivered into BPAEC via electroporation, and cells were plated at confluence. Immunoblot analysis for N-cadherin, VE-cadherin, and β-actin was carried out 48 hours after plating. Densitometric analysis: Values for N-cadherin and VE-cadherin are normalized to β-actin and are expressed relative to luciferase control (mean ± SEM, n=8; for N-cad P<0.0001, VE-cad P=0.0321 using single sample t-test. N siRNA: individual sequence targeting bovine N-cadherin purchased from Ambion.) * denotes significance (p<0.05) as determined using single sample t-test. (B) Confluent BPAEC monolayers were infected with either adenovirus containing GFP or one of two doses of flag epitope-tagged N-cadherin (AdN-cad-flag). After 48 hours, cells were lysed, and N-cadherin and VE-cadherin were detected in lysates by immunoblot analysis. β-actin was used as a loading control (bottom panels). (C) Top: Confluent BPAEC monolayers were infected with either GFP or one of three doses of VE-cadherin-flag adenovirus (AdVE-cad-flag) and immunoblotted for N-cadherin and VE-cadherin. Right – densitometric analysis: Densitometric values for cadherins are normalized to β-actin expressed relative to control GFP infection (mean ± SEM, n=3; for AdN-cad-flag, VE-cad P=0.0197 by one way ANOVA; for AdVE-cad-flag, N-cad P<0.0001 by one way ANOVA). Bottom: Following the 48 hour infection with AdVE-cad-flag, immunofluorescence microscopy was performed on BPAEC using an antibody to N-cadherin followed by a fluorescent-conjugated secondary antibody. Note the decrease in N-cadherin staining intensity with increasing dose of AdVE-cad-flag (left to right). Bar = 50 µm.