Fig. 1.

SPV122 potentiates MAPKi effects on M14 BRAF-mutant melanoma cells. a M14 melanoma cells have been exposed to encorafenib (BRAFi) starting from 5 μM and then diluted 1:2 for 10 times in the presence or not of SPV122 at fixed dose of 1.25 μM to measure cell viability through MTT assay after 72 h. b Crystal violet staining and O.D. at 595 nM reading by spectrometer assessed the growth inhibitory effects of encorafenib (BRAFi, 150 nM) and MEK162 (MEKi, 75 nM) in the presence or not of SPV122 (1.25 μM) for 72 h. The same drugs alone or in combination have been tested for apoptosis induction (c) and cell cycle (d) after 48 h of exposure. e M14 cells have been treated with the different drugs as previously described and total protein extracts have been subjected to Western Blot analysis to measure the expression levels of the indicated molecular effectors. f The same cells have been exposed two times a week with 250 nM of a BRAFi and then stained with Crystal violet (day 0). The remaining plates were treated with encorafenib in the presence or not of SPV122 (1.25 μM) and then stained after 3, 10, 20 and 30 days (left part). Quantification of data has been obtained as described above (right part)