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Figure 5


Fig. 5. Distributions of ZO-1 and perijunctional F-actin are altered by tMLCK expression. (A) ZO-1 profiles in monolayers not expressing tMLCK are smooth and arc-like. Only occasional mild undulations are present (arrow). (B) tMLCK expression causes ZO-1 profiles to be reorganized into irregularly undulating patterns (image shown is 4 hours after induction). These undulations are present throughout the monolayer (arrows). In addition, occasional cells are extremely affected (asterisk). Although relatively uncommon in induced cultures, such cells were never seen in monolayers without tMLCK expression. (C) Quantitative analysis of ZO-1 profile length shows that it is increased significantly (n=40 per condition; P<0.001) by tMLCK expression. MLCK inhibition using 250 µM PIK (Zolotarevsky et al., 2002) decreased profile length to levels observed in cells not expressing tMLCK (n=40 per condition; P>0.1). (D) Smooth F-actin and ZO-1 profiles are reorganized into irregular undulations 8 hours after tMLCK induction. Addition of 250 µM PIK (Zolotarevsky et al., 2002) during this tMLCK induction prevented the formation of undulations in both F-actin and ZO-1 distributions. Representative images are shown. Bars, 10 µm.





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