A, KRAS WT cancer cells (MNK45, STKM2, RKO) were infected with control lentivirus or lentiviral particles expressing KRAS-G12D. Western blot analysis data showed overexpression of KRAS-G12D significantly increased the protein levels of AURKA, p-RPS6KB1 (T389), p-MEK, and p-MTOR. B, KRAS-driven cancer cells (SW480, SNU-1, ESO26) were infected with control lentivirus or lentivirus-mediated expression of shRNA specific to KRAS, Western blot analysis data showed knockdown of KRAS decreased protein expression of AURKA, p-RPS6KB1 (T389), and p-MEK. C, KRAS WT cancer cells (STKM2, RKO) were infected as described in B. Western blot analysis data showed knockdown of KRAS decreased protein expression of p-MEK, without altering expression of AURKA and p-RPS6KB1 (T389). D-E, STKM2 and RKO cells were infected as described in A, then treated with alisertib for 3 days (D) or cells were transfected siAURKA for 48 hrs (E). Western blot analysis data showed downregulation of p-RPS6KB1 (T389). Representative blot is from one of two independent experiments. The relative density of bands are shown under immunoblot after normalization to the levels of actin). Quantification of Western blot data is included in Supplementary Figure S1 E to I. Black arrows indicate the molecular weight of proteins.