Figure 3.

Unc-51-like kinase 1 (ULK1) is a bona fide transcriptional target of p53. (a) Schematic representation of the putative p53 binding site of ULK1. (b). Electrophoretic mobility shift assay was carried out using biotinylated DNA oligos containing either the duplicated putative wild-type (wt) p53 binding sites (biotin-p53BS, lanes 1 and 3–5) or mutant (mt) p53 binding site (lane 2) in ULK1 gene as described in Materials and Methods. Wt (lane 4) or mt (lane 5) p53 binding sites of ULK1 were used for the competition assay. (The bands labeled with asterisk indicates nonspecific DNA complexes.) (c) Luciferase reporter assay was performed as described in Materials and Methods. Result for each condition was normalized to the firefly/Renilla luciferase activity ratio of empty control (lane 1). Ctr: empty pcDNA3.1 vector. Y axis is depicted as log scale. These data are representative of at least two independent experiments