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. Author manuscript; available in PMC: 2017 Jan 12.
Published in final edited form as: Oncogene. 2016 Jul 11;36(6):787–796. doi: 10.1038/onc.2016.247

Figure 4. FBW7 mutations are enriched in in CRC cells with acquired regorafenib resistance.

Figure 4

(A) Crystal violet staining of indicated parental and regorafenib-resistant (-R) CRC cells plated at equal density in triplicate after regorafenib (40 μM) treatment for 72 hr. (B) MTS analysis of cell viability of indicated parental (black) and regorafenib-resistant (-R) (red) CRC cells treated with increasing concentrations of regorafenib for 72 hr. Results were expressed as means ± s.d. of three independent experiments. (C) Sequencing of FBW7 genomic region from indicated regorafenib-resistant CRC cell lines highlighting the identified mutations. (D) FBW7 c.1513C>T and c.1393C>T mutations were analyzed by allele-specific PCR in HCT116, Lim1215, and Lim2405 cells after 1-4 rounds (R1-R4) of regorafenib selection. Genomic DNA from LoVo and CCK-81 cells spiked into FBW7-WT HEK293 cells at indicated ratios were used as controls for c.1513C>T and c.1393C>T mutations, respectively.