Fig. S8.

Formation of large aggregates in a hanging-drop assay is calcium dependent. (Top) Bright-field images showing aggregation of MDCK cells at 0- to 5-h time points in “normal” (1.8 mM) and low Ca²⁺ (5 µM) media. (Scale bar, 50 µm.) (Bottom) Quantification of hanging-drop assays in normal and low Ca²⁺ media. Cells were binned into cluster classes: 1–10 cells, 11–20 cells, 21–50 cells, 51–100 cells, or >100 cells. The percentage of cells in each cluster size for each time point is indicated. The data shown are averages from three independent experiments and error bars represent SEM.