ZO KD elevates afadin at the ZA. (A) Control and ZO KD cells cocultured. ZO-1 marks control cells. Junctional afadin increases after ZO KD. (B) Line scans from cocultures (10/image, three images). Error bars are SDs. Mean afadin intensity at BCJs is higher after ZO KD. au, arbitrary unit. (C) Afadin is enriched at TCJs (arrow) and MCJs (arrowheads). (C′) Spectrum display of image intensity. (D–G) Junctional ZO-1 or Ecad unchanged by afadin single KD.