Supplementary Figures from Tankyrase Inhibitors Target Colorectal Cancer Stem Cells via AXIN-Dependent Downregulation of c-KIT Tyrosine Kinase

Abstract

<p>Supplementary Figure 1 shows morphology, growth and plasticity of the CD44-positive and -negative COLO-320DM cells. Supplementary Figure 2 shows that tankyrase inhibitors preferentially target CD44-positive COLO-320DM cells. Supplementary Figure 3 shows that tankyrase inhibitors decrease the CD44-positive cell populations in colorectal cancer cells. Supplementary Figure 4 shows that tankyrase inhibitors upregulate AMOTL1 to comparable levels in CD44-positive and -negative COLO-320DM cells. Supplementary Figure 5 shows c-KIT expression in colorectal cancer cell lines. Supplementary Figure 6 shows that c-KIT depletion inhibits proliferation of CD44-positive COLO-320DM cells. Supplementary Figure 7 shows difficulties in detecting c-KIT phosphorylation in colorectal cancer cells under the normal growth conditions. Supplementary Figure 8 shows c-KIT phosphorylation induced by stem cell factor in CD44-positive COLO-320DM cells and its downregulation by tankyrase inhibitors. Supplementary Figure 9 shows that c-KIT overexpression strengthens resistance of DLD-1 cells to tankyrase inhibitors. Supplementary Figure 10 shows that G007-LK does not significantly destabilize c-KIT protein in COLO-320DM cells. Supplementary Figure 11 shows negative effect of imatinib on CD44-positive cell populations in COLO-320DM and DLD-1 cells. Supplementary Figure 12 shows that AXIN2 knockdown blocks c-KIT downregulation in G007-LK-treated colorectal cancer cells. Supplementary Figure 13 shows that G007-LK does not affect the stability of c-KIT mRNA in COLO-320DM and DLD-1 cells. Supplementary Figure 14 shows that tankyrase inhibitors downregulate c-KIT promoter activity in an AXIN2-dependent manner in DLD-1 cells. Supplementary Figure 15 shows that tankyrase inhibitors do not significantly downregulate SP1 protein levels in COLO-320DM and DLD-1 cells. Supplementary Figure 16 shows that tankyrase inhibitors downregulate JNK phosphorylation levels in CD44-positive COLO-320DM cells. Supplementary Figure 17 shows that G007-LK upregulates AXIN2 and downregulates c-KIT in vivo. Supplementary Figure 18 shows that tankyrase inhibitors do not enhance the therapeutic efficacy of irinotecan on DLD-1 xenograft tumors in vivo.</p>