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Fig. 2 Cell viability after treatment with cGMP analogs in melanoma spheroids and human melanocytes. (a) Dose?response relationships of melanoma cell spheroids viability after exposure to different doses of PA4 and PA5 for 24 h. Values of NT cells were set as 100% of cell viability. (b) Cell viability in human metastatic melanoma spheroids treated with PA5 (250 nM) for 24 h. Values of NT cells were set as 100% of cell viability. (c) Viability time course of melanoma cell line?derived spheroids treated with PA5 (250 nM). Cell viabilities at each time point were expressed as a percentage compared with that of time 0. (d) PA5 toxicity was evaluated in human primary melanocytes by MTT assay. Values of NT cells were set as 100% of cell viability. Data represent means ± SD of triplicate determinations. Student?s t-test was applied for statistical analyses of PA5-treated samples compared with NT. ??P < 0.01; ?0.01 < P < 0.05. h, hour; NT, not treated; PA, protein kinase G activator.