Figure 6.

Constitutively active ezrin rescues apical actin distribution in knockout cells. (A) Jeg-3–knockout cells expressing empty pCDH vector control, wild-type ezrin, or T567 phosphomimetic were stained for ezrin and actin. Maximum-intensity projections and vertical cross-sections (expanded fourfold for clarity) are shown. (B) Percentage of cells expressing microvilli-like structures. Bars represent mean ± SEM; n = 3. (C) Comparison of ZO-1 and actin between basolateral and apical confocal slices between Jeg-3–knockout cells expressing phosphomimetic ezrin-T567D. Apical Z-slices were determined by the presence of ZO-1. Yellow boxes indicate the region of the image that was expanded on the right. (D) Quantification of tight junction tortuosity using ZO-1 staining. n ≥ 10 cells per condition. Center lines show mean ± SEM. (E) The ratios of relative mean actin intensity values per cell between apical and basolateral cross-sections. n ≥ 15 cells per condition. Bars show mean ± SEM. P values were calculated against wild type with Welch’s t test (**, P ≤ 0.01; ***, P ≤ 0.001). Scale bars, 10 µm unless otherwise noted in magnified images.