Fig. 4

c-Myb binding to Sema5A promoter. a Western blot analysis of Bcl-2 and c-Myb expression and b qRT-PCR of c-Myb expression in A375SM-SC1 human melanoma control (empty) and Bcl-2 overexpressing (bcl-2) clones. mRNA levels were normalized using β-actin. Values are expressed as means of ratio ± SEM where ratio was calculated considering Bcl-2-overexpressing versus control clones. c c-Myb and d H3acPAN recruitment on Sema5A and Cyclin B1 (CCNB1) promoters by ChIP analysis. Values of each immunoprecipitated sample are expressed as percentage relative to their respective input and by subtracting the values obtained in the negative controls (no antibody).Values are expressed as ratio ± SEM where ratio was calculated considering Bcl-2-overexpressing versus control clones. *p-values < 0.05. e Western blot analysis of Sema5A and c-Myb expression in A375 and BV173 cells transduced with lentiviral vector for inducible c-Myb silencing. a, e, β-actin or HSP70/72 expression was evaluated to confirm equivalent transfer and loading. b, c, d, Experiments were performed at least three times (three technical replicates). *p-values < 0.05