Figure. 3. Cell lines harboring an A146T KRAS mutation demonstrate elevated RAS-GTP expression and KRAS-dependence.
A. HEK-293FT cells were transfected with KRAS mutants and GTP-bound RAS was measured by immuno-precipitating active RAS with recombinant Ras binding domain (RBD) of RAF. B. Colorectal cancer cell lines expressing A146T KRAS demonstrate elevated RAS-GTP expression as compared to cell lines with V600E BRAF mutation or those wild-type for both KRAS and BRAF. C. Knockdown of KRAS expression by siRNA in LS1034 cells (homozygous A146T KRAS mutant) inhibited colony formation.