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. 2020 Mar 4;21(4):e49269. doi: 10.15252/embr.201949269

Figure 2. WDR63 is a transcriptional target of p53.

Figure 2

  • A
    WDR63 mRNA and protein levels in A549, H292, and HCT116 cells transduced with empty vector (PCDH) or PCDH‐p53. Data shown are mean ± SD; n = 3 independent experiments. **P < 0.01; ***P < 0.001; two‐tailed Student's t‐test.
  • B
    WDR63 mRNA and protein levels in A549, H292, and HCT116 cells transduced with control (shctrl) or p53 (shp53) shRNA. Data shown are mean ± SD; n = 3 independent experiments. ***P < 0.001; two‐tailed Student's t‐test.
  • C
    A549 cells expressing control or p53 shRNA were treated with doxorubicin (Dox, 1 μg/ml) for the indicated periods of time. WDR63 mRNA and protein levels were then examined. Data shown are mean ± SD; n = 3 independent experiments. ***P < 0.001; two‐tailed Student's t‐test.
  • D
    WDR63 mRNA levels in H1299 cells transfected with empty vector (−), wild‐type p53, or the indicated p53 mutants. Data shown are mean ± SD; n = 3 independent experiments. ***P < 0.001; two‐tailed Student's t‐test.
  • E
    Schematic illustration of the first two exons of WDR63 gene. BS1 and BS2 represent 2 putative p53 binding sites predicted by the JASPAR database. Black bars represent p53 binding sites identified in the indicated p53 ChIP‐seq datasets. The pGL3‐based wild‐type and mutant reporter constructs used for luciferase assay are also shown.
  • F
    Lysates from A549 cells were subjected to ChIP assay using control IgG or anti‐p53 antibody. ChIP products were amplified by real‐time PCR. Data shown are mean ± SD; n = 3 independent experiments. ***P < 0.001; two‐tailed Student's t‐test.
  • G
    A549 cells were transfected with empty vector (ctrl) or Flag‐p53 plus the indicated reporter constructs. Twenty‐four hours later, reporter activity was measured. Data shown are mean ± SD; n = 3 independent experiments. **P < 0.01; ***P < 0.001; two‐tailed Student's t‐test.
  • H
    A549 cells expressing control or p53 shRNA were transfected with pGL3‐BS1 and Renilla luciferase plasmids. Twenty‐four hours later, cells were treated with or without Nutlin (10 μM) for another 12 h. Reporter activity was then measured. Data shown are mean ± SD; n = 3 independent experiments. **P < 0.01; two‐tailed Student's t‐test.