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Fig. 2 Novel p53 regulators activate or suppress p53 expression activity in mammalian cells. p53-positive regulators and control HIF1α (A) or negative regulators and control HDM2 (B) were transfected with FuGENE 6 into isogeneic HCT116 p53+/+ and p53-/- cells with the p53 reporter and a plasmid overexpressing β-galactosidase (β-gal). After 48 h incubation, cells were lysed, and luciferase activity was normalized based on β-gal activity. (C) Cell lysate from HCT116 cells transfected with p53 regulators were probed with antibodies against p53. (D) Reporter plasmid p53-1-luc was transfected into HCT116 p53-/- cells with p53 activators and β–gal plasmid. After incubating for 48 h, cells were lysed, and luciferase activity was normalized based on β-gal activity.