Figure 4

de novo CD133 induction by TGF-β correlates with overexpression of DNMT3 genes. a. the experiment in Figure 3A was repeated after MACS-sorting to enrich in CD133 negative cells, as depicted in the upper panel. Levels of CD133 expression were close to 0%, as shown in the upper histograms for both, Huh7 and HepG2 cells. Mean from three replicates is shown in the lower panels. (*) indicates P value < 0.05 relative to control conditions. b. sphere formation assays were performed in non-attachment plates, after exposure to TGF-β during 4 days. Spheres were counted after 6 days of growth in hepatosphere medium w/o TGF-β. c. Huh7 (left panels) and HepG2 (right panel) cells were treated as in (a), RNA was extracted and qRT-PCR was performed for genes involved in DNA methylation or demethylation. Expression was normalized to housekeeping gene GAPDH. (*) indicates P value < 0.05 relative to non-treated cells at the corresponding time point.