Fig. 5. ZRP-1-depleted cells fail to form N-cadherin-based cell-cell adhesions. (A) Actin dynamics at cell-cell contacts. Control cells (lower panel) and ZRP-1-depleted cells (upper panel) were transfected with EGFP-actin expression vector as described in Fig. 4. Cell-cell contact regions were then imaged at 1-minute intervals. Arrowheads indicate parallel actin filaments that connect neighboring cells. (B) ZRP-1-depleted cells transfected with EGFP-actin were imaged at 1-minute intervals. Red and yellow dashed lines outline two moving cells. (C,D) Three hours after switching the Ca²⁺ concentration of cell growth medium from low to high, both control cells (upper panels in C,D) and ZRP-1-depleted cells (lower panels in C,D) were fixed and stained with antibodies against ZRP-1 and phalloidin (C) or N-cadherin (D). Merged images are shown on the right of C and D. Bars, 15 µm. (E,F) Percentages of cell-cell adhesion with aligned actin filaments and N-cadherin signals. n indicates the number of cell-cell adhesions examined.