Fig 5.
MCV LT induces p53S15 phosphorylation through ATR/Chk1 pathway. (A) U2OS cells transfected with pcDNA4C (Vector) or pcDNA4C encoding the indicated Xpress-tagged MCV LT molecules were stained with Xpress (green) and pp53S15 (red) antibodies at 36 h posttransfection. The cells were counterstained with DAPI. Bar, 10 μm. (B) U2OS cells were transfected as in panel A. At 36 h posttransfection, cells were lysed and immunoblotted with the indicated antibodies. (C) U2OS cells transfected with pcDNA4C (Vec) or pcDNA4C-LT1-817 were treated with dimethyl sulfoxide (DMSO) or 10, 20, or 30 μM NU6027 for 24 h. Cells were lysed and immunoblotted with the indicated antibodies. (D) U2OS cells were transfected with control (CTRL) or ATR siRNA. At 36 h posttransfection, cells were transfected with pcDNA4C (V) or pcDNA4C-LT1-817 (LT). Cells were lysed and immunoblotted at 72 h after siRNA transfection with the indicated antibodies. (E) U2OS cells transfected with pcDNA4C (Vec) or pcDNA4C-LT1-817 (MCV LT) were treated with DMSO or 5 nM AZD7762 for 24 h. Cells were lysed and immunoblotted with the indicated antibodies. All experiments were repeated three times with consistent results.