Fig. 6: Knockdown of E-cadherin contributes to epithelial dysfunction.
From: Loss of E-cadherin is causal to pathologic changes in chronic lung disease
To knock down E-cadherin in airways, mice tracheal epithelial cells (mTECs) from Cdh1fl/fl mice cultured at the air-liquid interface (ALI) were transfected with Ad5CMVCre-eGFP (Cre) at 2 × 109 pfu and these were compared to Ad-5CMVeGFP (Ctrl). mTECs transfected with Cre show reduction in E-cadherin as assessed by a mRNA expression of Cdh1 (encodes for E-cadherin), and b western blotting (representative image – left panel, and quantification – right panel). c Epithelial resistance was decreased, and d cellular velocity was increased in mTECs with E-cadherin knockdown. Data is expressed as median bars and generated of cells derived from 12 mice, 4 to 12 inserts. Normal human bronchial epithelial cells at ALI (normal controls) were transfected with Ad-GFP-U6-h-CFL1-shRNA (shCDH1) at 1.5 × 109 pfu to knock down E-cadherin and these were compared to control adenovirus (Ad-GFP-U6-shRNA, GFP). Normal control cells transfected with shCDH1 show reduced e mRNA of CDH1 (encodes for E-cadherin) and f protein expression of E-cadherin (representative blot—left panel, and quantified blot—right panel). Assessment of the epithelial barrier function indicates that g monolayer resistance was decreased, h a trend towards increased permeability, and i cellular velocity was increased in control cells with knockdown of E-cadherin. Data is expressed as median bars and representative of 5 to 10 inserts per condition. Mann-Whitney test was performed and P < 0.05 was considered statistically significant.
