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Analysis of Sphere Formation in Cell Cultures Induced by EGF/bFGF and HGF Treatments

This study presents supplementary figures detailing the effects of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and hepatocyte growth factor (HGF) on the sphere formation in various cell lines. We analyze the proportion of wells without sphere formation under different treatment conditions, highlighting differences when using specific inhibitors like PHA665752, SU11274, and SGX523. Additionally, we examine expression levels of WNT target genes in cell cultures and assess the impact of growth factors on cellular behavior.

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Analysis of Sphere Formation in Cell Cultures Induced by EGF/bFGF and HGF Treatments

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  1. Supplementary Figure 1 A. 827 + EGF/bFGF 827 + EGF/bFGF 827 + EGF/bFGF 0 PHA665752 SU11274 SGX523 -1 Proportion of wells w/o sphere (log10) -2 -3 -4 0 50 100 150 200 0 10 20 30 40 50 0 5 10 15 20 827 + HGF 827 + HGF 827 + HGF 0 PHA665752 SU11274 SGX523 -1 Proportion of wells w/o sphere (log10) -2 -3 -4 0 50 100 150 200 0 20 40 60 80 100 0 20 40 60 80 100 number of plated cells B. 131 + EGF/bFGF 131 + EGF/bFGF 131 + EGF/bFGF 0 PHA665752 SU11274 SGX523 -1 Proportion of wells w/o sphere (log10) -2 -3 -4 0 100 200 300 400 500 20 40 60 80 100 0 0 50 100 150 200 131 + HGF 131 + HGF 131 + HGF 0 PHA665752 SU11274 SGX523 -1 Proportion of wells w/o sphere (log10) -2 -3 -4 0 50 100 150 200 0 50 100 150 200 0 50 100 150 200 number of plated cells

  2. Supplementary Figure 2 464T +EGF/bFGF 464T +EGF/bFGF -SU11274 464T +EGF/bFGF +SU11274 2.08% 31.8% 25.4% CD133-APC Isotype-APC 464T +HGF 464T +HGF -SU11274 464T +HGF +SU11274 Isotype-APC 2.83% CD133-APC 9.00% 7.53%

  3. Supplementary Figure 3 GFP ABC Merge Merge + DAPI Field 1 GFP ABC Merge Merge + DAPI Field 2 GFP ABC Merge Merge + DAPI Field 3

  4. Supplementary Figure 4 mCMV backbone reporter mCMV-TCF/LEF reporter 464T Relative Luciferase Unit Wnt3a (ng/ml) 0 25 50

  5. Supplementary table 1 Expression levels of WNT target genes in METhigh/+ cells

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