Figure 4

Wild-type and mutant p53s regulate miR-520G and melanoma antigen expression without inducing considerable cell death. (a) A map of CDKN1A gene’s p53 binding palindromic boxes-1 (green font/box) and 2 (orange font/box) [CP box] within C19MC (grey box) and surrounding landscapes. 6-mer-to-10-mer palindromic sequences were mapped to a 200 kb region enclosing C19MC. (b) A map of GADD45 gene’s p53 binding palindromic box (6-mer palindrome = green font/box), (8-mer palindrome = pink font/box) and (7-mer palindrome = orange font/box with 1 nucleotide match to 8-mer palindrome blue font) [GP box] within C19MC (grey box) and surrounding landscapes. (c) Transient transfection of p53-WT or mutant p53s does not induce considerable mitochondrial depolarization in Hep3B cells as evaluated by JC1 and Hoechst-33342 staining. The microscopy images were pseudo-colored as indicated. Purple corresponds to JC1 staining of depolarized mitochondria (MOMP). Also see Fig. 6D-E for more details. (d) Transient transfection of p53-WT or mutant p53s induce C19MC miRNA-520G expression in Hep3B cells as evaluated by qRT-PCR and is associated with induction/promotion of MAGEA mRNAs as evaluated by RT-PCR (panel-e).