Fig. 6. The effect of TGFß on endoderm. Day 13 EBs were treated with various concentrations of TGFß1 for 5 days. (A) The expression of AFP was assessed by qRT-PCR and normalized to GAPDH. Results are presented relative to expression in untreated samples. (B) AFP protein expression in untreated and TGFß-treated (1 ng/ml) EBs was compared by western blotting. An anti-ß-actin antibody was used as a loading control. The expression of AFP (C,D) in control and treated samples was examined by immunofluorescence staining. DAPI-stained nuclei are also shown (E,F). Day 3 EBs were treated with various concentrations of TGFß1 for 5 days. The RNA level of (G) AFP and (H) TTR and (J) THBD was measured relative to GAPDH. Results are presented relative to expression in untreated samples. (I) Endo A protein expression in untreated and TGFß-treated (1 ng/ml) EBs was compared by western blotting.