Dock1 and Elmo2 are essential for early E-cadherin recruitment to cell–cell contacts. (A) Basal focal plane of Lyn-GFP expression in cells treated with indicated siRNAs and fixed 90 min after plating on collagen. (B) Box plot quantification of Lyn-GFP fluorescence intensity for an equal surface area in indicated siRNA-treated cells (n = 40 for each condition). Whiskers show minimum and maximum values, horizontal lines show medians, and boxes show 1st and 3rd quartiles. P-values were determined by unpaired t test for the indicated samples. (C) E-cadherin immunofluorescence in indicated siRNA-treated cells at different times after calcium addition to induce cell–cell adhesion. (D) Box plot quantification of the ratio of E-cadherin fluorescence intensity at a region of cell–cell contact normalized to the intensity of an equal region of the cytoplasm underlying the contact (n = 100 for each condition). P-values were determined by unpaired t test for the indicated samples. (E) Quantification of hanging drop assays for the indicated siRNAs in which the cells were binned into cluster classes: 1–10, 11–20, 21–50, 51–100, or >100 cells. The percentage of cells in each category is shown for each time point. The data shown are from a representative experiment from three repeats in which ∼5 × 10⁴ cells were analyzed for each time point. (F) Montages of individual frames from Video 1 of cells expressing E-cadherin–GFP treated with scramble or Elmo2 siRNA. Yellow arrows indicate the boundary of the cell–cell contact. Bars, 5 µm.