Absence of activated ERM proteins alters cell–cell junctions. (A) Comparison of ZO-1 and actin between apical and basolateral confocal slices. Apical Z-slices were determined by the presence of ZO-1. Scale bars, 10 µm unless otherwise noted in magnified images (2 µm). (B) Intensity of actin across junctions highlighted in blue in A. (C) Quantification of tight junction tortuosity using ZO-1 staining. Each point represents the tortuosity from one tight junction intersection to the next intersection. Center lines represent mean ± SEM; n ≥ 10 cells per condition. (D) The ratios of relative mean actin intensity values per cell between apical and basolateral cross-sections. Bars represent mean ± SEM; n ≥ 10 cells per condition. P values were calculated with Welch’s t test (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001). A.U., arbitrary units.