Fig. 6. Maturation of cell-cell adhesions. (A,B) Ca²⁺-switch assay was performed with CTRL siRNA (A) or WAVE2 siRNA (B) treated confluent cell layers. Confluent cell layers were treated with 4 mM EGTA and then incubated with Ca²⁺-containing medium for the indicated times. Cell layers were fixed and stained for ß-catenin and actin filaments (F-actin). Arrows indicate disorganized cell-cell adhesions. Bar, 10 µm. (C) Cell-cell contacts were classified into three categories on the basis of the ß-catenin staining pattern. Categories were thick, strong signals (thick); thin, weak signals (thin); and broad or no signal (disorganized). All data are mean ± s.d. Three independent experiments were performed. (D,E) Vertical sections of CTRL siRNAi (D) or WAVE2 siRNA (E) treated cells in the Ca²⁺-switch assays. Brackets indicate the width of cell-cell adhesions. Bar, 7.5 µm.