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. 2021 Feb 16;49(5):2759–2776. doi: 10.1093/nar/gkab084

Figure 4.

Figure 4.

DAZAP2 interacts with p53. (A) A fraction of DAZAP2 accumulates in the nucleus upon DNA damage. HCT116 cells were treated with DMSO or 50 μM 5-FU for 24 h. Cells were fractionated into nuclear and cytoplasmic fractions, and equal total protein amounts of all fractions were analyzed by immunoblotting. WCL = whole cell lysate. Cyto = Cytoplasm. n = 3. (B) Interaction of endogenous DAZAP2 and p53. Lysates from untreated or Adriamycin-treated HepG2 cells in the presence of MG-132 were subjected to immunoprecipitation with control or p53 antibodies. Lysates and immunoprecipitates were analyzed by immunoblotting. n = 2. (C, D) In vitro interaction of DAZAP2 and p53. (C) GST pull-down assays were performed using in vitro translated 35S-labeled p53 and bacterially expressed GST-DAZAP2 or GST as a control. n = 3. (D) GST pull-down assays were performed using in vitro translated 35S-labeled DAZAP2 and bacterially expressed GST-p53 or GST as a control. n = 3. GST pulldown assays were analyzed by SDS-PAGE, Coomassie Brilliant Blue (CBB) staining and autoradiography. Representative experiments are shown.