EPLIN is specifically important for ZA formation. (A) Double-immunostaining for E-cadherin and vinculin (left) or EPLIN (right). Images are focused on ZAs in inner cells and pAJs in peripheral cells. Vinculin colocalizes with E-cadherin in both lAJs and pAJs, whereas EPLIN is not detectable in pAJs. Arrows point to typical E-cadherin signals, associated with vinculin or not associated with EPLIN. (B) The effects of EPLIN depletion on junctional organization. Cells located in the inner portion of the colony were double-immunostained for F-actin and myosin IIA. Note that the junction morphology in EPLIN-depleted (siEPLIN) cells is similar to that observed in the peripheral cells of control colonies (see Fig. 1 C). Bars, 10 µm.