Figure 4

EpCAM patterning in differentiating ESC. (a) EpCAM expression during 3D-differentiation of ESC is depicted through immunohistochemistry staining in E14TG2α EB (d3–6; scale bars represent 50 µm). (b) Representative EpCAM staining in d14 and d21 EB showing basolateral staining in differentiated epithelia. (c) EpCAM, Foxa2 (endoderm), vimentin (mesoderm) and nestin (ectoderm) expression in consecutive sections of EB at d4 (scale bars represent 100 µm). (d) EpCAM and vimentin expression in consecutive sections of EB at d4 (200x and 400x magnification; scale bars represent 50 and 20 µm). (e) EpCAM and vimentin expression in EB at day 4 (upper) and 5 (lower) depicted as double-immunofluorescence staining (EpCAM: green; vimentin: red). Left panels comprise full overviews of EBs; right panels represent magnifications of the area demarked in the doted squares. Shown are representative pictures from ≥3 independent experiments with multiple EB each (scale bar represents 50 µm). Staining of the indicated markers is depicted in brown. Cytoplasm and nucleus were counterstained with H&E, except for immunofluorescence staining in (e).