Fig. 2: TRMP is a direct transcriptional target of p53.

a H1299 cells were infected with lentiviruses expressing wild-type p53 or the indicated p53 mutant forms. Forty-eight hours later, total RNA was subjected to real-time RT-PCR analysis to examine TRMP levels. Data shown are mean ± SD of three independent experiments. Cell lysates were also analyzed by western blotting. b Schematic illustration of two putative p53 binding sites (P1 and P2) in TRMP gene promoter. c Shown are the pGL3-based wild-type and mutant reporter constructs used for luciferase assay. d H1299 cells were co-transfected with either control vector, Flag-p53, or together with the reporter constructs in the indicated combination. Twenty-four hours after transfection, the reporter activity was measured and plotted after normalizing with respect to Renilla luciferase activity. Data shown are mean ± SD of three independent experiments. *** and n.s. indicate p < 0.001 and no significance, respectively. e U2OS cells expressing either control shRNA or p53 shRNA were co-transfected with the indicated reporter constructs and Renilla luciferase plasmid. Twenty-four hours after transfection, the reporter activity was measured and plotted after normalizing with respect to Renilla luciferase activity. Data shown are mean ± SD of three independent experiments. ** and n.s. indicate p < 0.01 and no significance, respectively. f Lysates from U2OS cells were subjected to ChIP assay using anti-p53 antibody or an isotope-matched control IgG. ChIP products were amplified by real-time RT-PCR with the indicated pairs of primers. Data shown are mean ± SD of three independent experiments. ** and n.s. indicate p < 0.01 and no significance, respectively