Figure 3
From: Three-dimensional Organotypic Culture Models of Human Hepatocellular Carcinoma
(A) Representative images of Huh1 spheres at Day 7 with no treatment (top panels) and 2.5 ng/ml of TGF-β added at day 5 of spheroid culture (bottom panels). Shown are spheres immunostained for E-cadherin (a,f; green), EpCAM (b,g; red) and merged (c,h). Other representative spheres immunostained for vimentin (d,i; red) and merged (e,j) are also shown. Scale bars are 50 μm. (B) The expression level of E-cadherin, vimentin, phosphorylated Smad2 and EpCAM in Huh1 spheres treated with 5 ng/ml TGF-β at day 3 of spheroid culture. (C) Relative number (black bar) and the acini proportion (grey bar) of Huh1 spheres in the presence of TGF-β are shown. Data are presented as the average of three independent analyses. (D) Representative images of round-shape cell spheres (top, left) and protruding cell spheres (top, right) are shown. The proportion of each sphere type following treatment with 5 ng/ml TGF-β combined with several kinds of TGF-β receptor kinase inhibitors are shown as a bar graph (bottom). Data are presented as the average of duplicate experiments. All error bars represent ±SEM. *p < 0.05.
