The sensitivity of MAP2K1 variants to BRAF and MEK inhibitors using the MANO method. A375 cells with MAP2K1 variants, GFP were treated with DMSO or BRAF inhibitors (vemurafenib, dabrafenib, and encorafenib) and MEKis (cobimetinib, trametinib, and binimetinib) at the indicated concentrations. Concentrations of the BRAF and MEK inhibitors were varied by 6 (0 to 10,000 nmol/L) and 12 steps (0 to 10,000 nmol/L), respectively. The relative viability of the treated cells with each drug versus DMSO-treated cells was measured, and the results are presented using a color-coded scale. Data are presented as the mean ± SD (n = 3). A, Combination therapy with cobimetinib alone (top) and vemurafenib 100 nmol/L in addition to cobimetinib (bottom). B, Combination therapy with trametinib alone (top) and dabrafenib 10 nmol/L in addition to trametinib (bottom). C, Combination therapy with binimetinib alone (top) and encorafenib 10 nmol/L in addition to trametinib (bottom).