Ectodermal and endodermal differentiation. Neuroglial differentiation was evidenced by morphology after 10 days of induction (A), expression of neuroectodermal-specific genes (B), and by immunofluorescence assay for MAP-2 (C) and GFAP (D). Calcium imaging assays for the change in [Ca²⁺]_i (E) and the ratio of change (F), as determined by fura-2 fluorescence, in the presence of high extracellular potassium. Hepatocytic differentiation was evidenced by morphology after 4 weeks of differentiation (G), expression of liver-specific genes (H), immunofluorescence assay for human albumin after 7 days of differentiation (I), and uptake of LDL after 6 weeks of differentiation (J). Original magnification, × 100 for all panels. Error bar in panel F indicates standard deviation.