Imaging of junctional and apical defects with myo-IIA localization and inhibition by blebbistatin. (A) Deconvolution immunofluorescence staining of myo-IIA shows a similar organization to myo-IIB (Fig. 4). Nonmuscle myo-IIA and myo-IIB are localized to aberrant apical contractile fibers. (B) Jeg-3 wild-type and knockout cells were treated with 25 µM blebbistatin for 30 min before immunofluorescence staining of ZO-1 and actin. Images are maximum-intensity projections of apical Z-slices as determined by ZO-1 signal. (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 ≥ 13. Scale bars, 5 µm in A, 10 µm in B.