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Fig. 4. (A) Effect of RA treatment on N-Myc expression in parental and nNOS overexpressing cells (clone 4 and pooled clones). Levels of N-Myc were evaluated by western blot. (B) N-Myc was quantified through its ratio to actin content of the samples. Bars are the mean±s.e.m. of four experiments. *, P<0.01 compared with control (Bonferroni's test after ANOVA). (C) Effect of RA treatment on N-Myc in clone 4 after the addition of the NOS inhibitor L-NAME (1-5 mM) and the guanylate cyclase inhibitor ODQ (50 µM) to the culture medium. (D) The negative regulation of N-Myc protein expression was replicated in parental cells using increasing concentrations of DETA NONOate (50-200 µM) and the cGMP analog 8Br-cGMP (250 µM).