Figure 5. Effects of Dabrafenib, AZ628, and Trametinib alone or in combination on H1666 cells.

H1666 Cells were incubated for 48 h with Dabrafenib (2.5 μM), AZ628 (2.5 μM), or Trametinib (25 nM) alone or in combination (Dabrafenib or AZ628 plus Trametinib). Whole cell lysates were subjected to Western blot analysis (A and B). H1666 cells were incubated for three days with Dabrafenib (2.5 μM), AZ628 (2.5 μM), or Trametinib (25 nM) alone or in combination (Dabrafenib or AZ628 plus Trametinib). Viability was measured, and relative viability was determined via normalization to the vehicle group (C). Means ± SEM are from two independent experiments, each performed in six replicates. All three inhibitors showed comparable effects as single agents, and combined RAF/MEK inhibitor treatments were more efficient. H1666 cells were incubated for three days with incremental doses of Dabrafenib and AZ628 and a constant Trametinib dose (25 nM). Viability was measured, and relative viability was determined via normalization to the vehicle group (D). Means ± SEM are from three independent experiments, each performed in four replicates. Cells were incubated for three days as in (D) and Caspase-3/7 activity was measured and normalized to the number of viable cells (E). Single agent treatments had no pro-apoptotic effects. AZ628 plus Trametinib showed a stronger pro-apoptotic effect than Dabrafenib plus Trametinib. Values are displayed as fold increase compared to the vehicle group. Means ± SEM are from two independent experiments, each performed in six replicates. ^*p ≤ 0.05, ^**p ≤ 0.01, ^***p ≤ 0.001.