Fig. 2

Sorting patterns in aggregates of wild-type and E-cadherin deficient ES cells. a Two representative cell sorting patterns: dispersed single cells from RW4 wild-type and E-cadherin-deficient 9j ES cells were mixed and allowed to aggregate for 2 days in suspension culture. The resulting cell aggregates were analyzed by histology, and 5 μm sections on glass slides were stained with E-cadherin and actin, countered stained with DAPI. Representative confocal images are shown for two main cell sorting patterns: either the wildtype (E-cadherin positive) cells were clustered in the center, surrounded by E-cadherin negative cells (upper panel); or the wildtype cells formed a shell on the surface, enveloping E-cadherin negative cells (lower panel). b Intermixed ES cells, either RW4 wildtype plus CFG37 (GFP-labeled WT), or the E-cadherin deficient 9j plus GFP-labeled WT, were allowed to aggregate in suspension culture for 2 days. Cryo-sections of the spheroids were analyzed by GFP epifluorescence, immunostaining of E-cadherin, and countered staining with DAPI. Representative examples are shown: one image from WT + WT-GFP, and three images from E-cadherin (−/−) + WT-GFP spheroids. Each sorting patterns ranked from 10 to 70% of the aggregates, variable in each independent experiment performed