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. 2012 Oct 14;2012:617085. doi: 10.1155/2012/617085

Figure 2.

Figure 2

Huh 7 side population cells possessed higher clonogenicity and expression of stem cell markers. (a) Cell sorting of Huh7 cells using Hoechst 33342. The R2 gate represented the SP cells (1.69% of total cells) and the R1 gate represented the non-SP cells. Both of them were collected for the subsequent experiments. (b) RT-PCR assay showed that the SP cells possessed higher expression of stem cell markers. (c) Clonogenic assay showing the clonogenicity of SP cells was markedly higher than that of non-SP cells. (d) Cell morphology of freshly sorted SP and non-SP cells at days 0, 3, and 9 after seeding. As shown in the photographs, the differences in the shapes of SP (small and round in shape) and non-SP cells (fibroblast-like) were remarkable. (e) Protein expression and nuclear location of β-catenin in SP and non-SP cells. The SP cells had much higher nuclear β-catenin than the non-SP cells. Similar phenomenon was also observed in the cytoplasmic β-catenin level. Scale bar, 50 μm. Similar results of clonogenicity and stemness genes expression were also obtained in HepG2 and Hep3B SP cells (data not shown). NSP: non-SP cells.