The high levels of β-catenin signaling do not up-regulate Nanog and Oct4 expression in both HONE1 and SLMT1 cells. A. Western blotting shows that both concentrations of 5 μM and 10 μM BIO treatment induce strong over-expression of β-catenin in treated cells, but 10 μM BIO treatment causes strong cytotoxic effects in cells and possible reduced expression of Axin2 in HONE1 cells. Compared to mock-treated HONE1 and MCH4.5-2TS cells, no obvious up-regulation of both Nanog and Oct4 expression is detected in β-catenin over-expressing cells, but E-cadherin is clearly up-regulated in both BIO-treated cell lines. B. SLMT1 cells have high levels of expression of β-catenin and do not respond to exogenous β-catenin signaling. RT-PCR analysis shows that HONE1 hybrid cells have an up-regulated expression of β-catenin and c-Myc compared to their parental HONE1 cells, but no up-regulated change of β-catenin (exons 3/5) and c-Myc is detected in SLMT1 and its hybrids, MCHs 3.11 and 3.22. C. Western blotting analyses do not detect changes of proteins β-catenin, Nanog, and Oct4 in SLMT1 hybrid cells, compared with their parental SLMT1 cells.