Fig 7.

Embryoid body culture and expression analysis of molecular markers. (A) Morphology of embryoid bodies that have been treated (A) or untreated (A') with TGF-β1 (10ng/ml) for 4 days. Arrows in A point to cavities induced by TGF-β1. (B-D) Gene expression of in TGF-β1 treated (B-D) or untreated (B'-D') embryoid bodies revealed by wholemount in situ hybridization. (E). RT-PCR analysis of gene expression. RNA was isolated at different time points from both TGF-β1 treated (T) and untreated (C) embryoid bodies. TGF-β1 was added 48 hours after starting the culture, so the third day counts as treatment day 1 (T1). The treatment was lasted for 4 days. (F) RNA was isolated from both wild type and SMAD4-/- embryoid bodies at different time points, from day 0 (ES), 3 (EB3), 6 (EB6), 9 (EB9) and 12 (EB12).