Figure 3. Viability and proliferation assays of double mutated cells.

(A) Viability assays of double mutated cells for different MAPK inhibitors and inhibitors from the PI3K-AKT pathway as well as a multi-RTK inhibitor. Single mutated control cell cultures are M000921 (BRAF^V600E) and M010817 (NRAS^Q61R), indicated in dotted lines. The double mutated cell cultures are indicated in solid lines. MM121224 (BRAF^V600E, NRAS^Q61K) derives from patient 1, MM140307 (BRAF^V600K, NRASG12A) derives from patient 5, MM140906 (BRAF^V600E, NRAS^Q61R) derives from patient 6, MM150423 (BRAF^V600E, NRAS^Q61R) derives from patient 4, M130903 (BRAF^V600E, NRAS^Q61H) derives from patient 7, MM150849 (BRAF^V600E, NRAS^Q61R) and MM150850 (BRAFV^600E, NRAS^Q61K) are both derived from patient 8. (B) Doubling time of double mutated cells under standard culturing conditions. Single mutated control cell cultures are indicated with stars. (C) Western blots showing pERK and ERK levels under MAPK inhibitor treatment in double mutated cell cultures. Single mutated control cell cultures are indicated with stars. (D) Westernblot showing pERK and pAKT levels under basic conditions (no treatment) in the different double mutated cells.