QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 8. Compromise of TJ integrity following knockdown and rescue of Bves in HCE cells. (A) Expression of Bves and ZO-1 at the cell periphery in parental HCE cells. (B) HCE-R cells stably expressing the `rescue' chicken Bves-FLAG construct exhibit labeling of the membrane by both FLAG and Bves antibodies. Note the intracellular accumulation of transfected chicken Bves. (C) Treatment with anti-human Bves MO results in a loss of Bves at the cell periphery, as compared to untreated (NT) and control MO. (D) TER measurements were performed on HCE and HCE-R. Parental HCE cells exhibited a TER value of approximately 390 /cm². Treatment with anti-human Bves morpholino resulted in a 40% decrease (P<0.05) in TER (asterisk), while control MO did not significantly alter the TER. The TER of HCE-R cells doubled to 775 /cm² and, following anti-human Bves MO (double asterisk), a drop of only 20% (P<0.05) in TER was observed. (E) ZO-1 immunofluorescence performed on Transwell inserts following TER measurements. Parental HCE cells and HCE-R cells were treated with control and anti-human Bves MO. Bves and ZO-1 expression is disrupted in HCE cells treated with anti-human Bves MO (top right panel). Morpholino treatment does not alter the expression pattern of ZO-1 in HCE-R cells (lower right panel). (F) Western blotting demonstrates that membrane ZO-1 and Bves increase with exogenous expression of Bves in HCE-R cells, but decrease following treatment with Bves-MO, as compared with the parental HCE line. (G) Membrane integrity of junction proteins occludin, E-cadherin and ß-catenin is disrupted following Bves MO treatment. Scale bars: 10 µm.

Return to article