Expression and localization of junctional proteins.

The junctional localization of various proteins involved in formation of adherens junction and tight junctions were determined by immunofluorescence staining using specific antibodies to VE-cadherin, N-cadherin, β-catenin, p120 catenin, and ZO-1 as detailed in Methods (A). The changes in the levels of these proteins were determined by Western blot analysis of cell lysates using specific antibodies (B). The quantitative assessment of the data is shown in (C). Please note a significant decrease in the levels of VE-cadherin and ZO-1 proteins in Cyp1b1-/- LSEC, while the levels of N-cadherin and p120 catenin were modestly increased (****P< 0.0001 and **P< 0.01; n = 3). These results are consistent with the intensity of staining seen in (A). These experiments were repeated with two isolations of LSEC with similar results.