Figure 6.

A. Sequential TER measurements of MDCK II cell monolayers that were treated with 10 mM MBCD for 2 h (white circles). The control monolayers (black circles) underwent medium changes at the same time that MBCD was either added or removed from the experimental monolayers. TER falls to near zero within 2 h of adding MBCD. Within 24 h of removing MBCD, TER returns to control levels. B. Occludin was immunoprecipitated from untreated monolayers or from monolayers incubated for 0.5 or 2 h with 10 mM MBCD and the immunoprecipitates were subjected to Western blotting. The Western blots were probed with anti-occludin, anti-phosphotyrosine or anti-phosphothreonine antibodies. C. Monolayers of MDCK II cells were treated with 10 mM MBCD without or with 100 μM Genistein (G) for 0, 0.5 or 2 h. Occludin was then immunoprecipitated and Western blots were probed using either anti-occludin (loading control) or anti-phosphotyrosine antibodies. TER data are the mean +/- SD of triplicate inserts. Data in Fig. 6A and B are representative of three independent experiments and the data in Fig 6C are from a single experiment.