As a biologically relevant test of CycIF we imaged BRAFV600E melanoma cells (Fig. 3, Supplementary Fig. 9), in which the therapeutically beneficial effects of Raf inhibition are opposed by counter-therapeutic upregulation of MAPK, Akt/mTOR and other compensatory progrowth pathways (so-called adaptive responses27). The gene discussed is AKT1; the disease is melanoma.