Supplementary Figures S1-S15 from Targeting the c-Met/FZD8 Signaling Axis Eliminates Patient-Derived Cancer Stem–like Cells in Head and Neck Squamous Carcinomas

Supplementary Figures S1-S15. Identification and enrichment of CSCs directly from clinical SCCHN samples ; qRT-PCR measurement of the c-Met mRNA level and western blot measurement of c-Met protein in HN-CSC#1 spheres, HN-CSC#2 spheres, and HN-CSC#3 spheres infected with lentiviral nonspecific shRNA (scramble) or c-Met-specific shRNAs ; Knockdown of c-Met inhibited the properties of HN-CSCs ; Dose-dependent effect of conventional chemotherapeutic agents (cisplatin, 5-Fu, and docetaxel) on the viability of tumor cells derived from patient HN-0-22, HN-0-29, or HN-0-46 ; Treatment with c-Met inhibitor PF-2341066 inhibited the formation of HN-CSC#2 spheres and HN-CSC#3 spheres in a dose-dependent manner ; Treatment with c-Met inhibitor SU11274 inhibited the formation of HN-CSC#1 spheres and HN-CSC#2 spheres in a dose-dependent manner ; The combination of c-Met inhibition and docetaxel treatment eliminates CSCs in patient-derived tumor xenografts ; Pharmacologic inhibition of c-Met enhances the therapeutic efficacy of conventional anti-cancer agent 5-Fu ; Western blot analysis shows that no anaplastic lymphoma kinase (ALK) expression was detected in seven SCCHN PDXs including HN-0-22, HN-0-29, and HN-0-46-derived xenografts ; Histologic confirmation of metastasis by H&E staining ; RT2 Profiler PCR Array and further qPCR validation ; Downregulation of FZD8-mediated Wnt/β-catenin signaling is a crucial step for the elimination of HN-CSCs caused by c-Met inhibition ; Quantification of the normalized photon flux, measured at weekly intervals following cells inoculation ; Western blot analyses ; C-Met functionally regulates FZD8 through an ERK/c-Fos cascade in HN-CSCs .