Fig. 4. Combination treatment with AKT and PAK inhibitors in BRAFV600E cell lines.
(A) Thyroid cancer cell lines were treated with increasing doses of AKT inhibitor (MK2206) alone for 72 hours. Cell viability was determined by WST-8 assay. The optical density of each treatment was normalized to the 0 μM control. Assays were conducted in triplicate with at least three biological replicates. Data are represented as means ± SD. (B) WST-8 cell viability assay of K1 and SW1736 after 72 hours of combined AKT (MK2206) and PAK (G-5555) inhibition. All dose combinations were normalized to the 0 μM G-5555 + 0 μM MK2206 control. Experiments were conducted in triplicate with ≥ three biological replicates. Data are represented as means ± SD. (C) Synergy tables representing MK2206/G-5555 combinations as per Zhao et al (Zhao et al. 2012) and as described in Fig. 3. (D) pPAK S144/S141, pAKT S473, and pGSK-3αβ S21/9 are shown at doses and timing selected for synergy and cell viability. GAPDH is loading control for each membrane above it. At least two biological replicates were performed per cell line with similar results.