Feedback from tissue-level contractility and myosin activity is required for myosin accumulation but not afadin recruitment. Control, ZO KD, or mixed culture projections of apical 3-µm sections. Some cells treated with DMSO (control; A, I, and K); 0.1 mM blebbistatin (J); or 50 µM Y27632 (L). (B and D–F) Small islands of control cells in ZO KD monolayers. Each cell type retained patterns of myosin (D), afadin (E), and vinculin (F) seen in unmixed monolayers (e.g., A). (C, G, and H) Small islands of ZO KD cells in control cell monolayers. ZO KD cells had reduced cortical myosin (G), whereas afadin remained elevated cortically but became less continuous (A vs. H′). (I and J) Blebbistatin fragmented cortical myosin and afadin (I vs. J). (K and L) ROCK inhibition also made afadin more discontinuous.