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. 2003 Sep;23(17):6174–6186. doi: 10.1128/MCB.23.17.6174-6186.2003

FIG. 1.

FIG. 1.

ZIPK colocalizes with Daxx in PODs. (A) HeLa cells grown on coverslips were transiently transfected with FLAG-ZIPK. After 36 h of transfection, cells were stimulated with 1,500 U of IFN-γ/ml or 1.0 μM As2O3 for 12 h. After fixation, cells were stained with an anti-FLAG antibody (M2) for the detection of ZIPK and DAPI for visualization of nuclei. Antibody detection was achieved with a FITC-conjugated anti-mouse antibody (green), followed by microscopy analysis. The bottom panels show the overlay result. (B) HeLa cells transiently transfected with FLAG-ZIPK were stimulated with IFN-γ and As2O3 for 12 h and stained with an anti-FLAG antibody, and the percentage of transfected cells with a speckled pattern was determined by counting a minimum of 200 transfected cells. Data represent means ± standard deviations (n = 3). (C and D) HeLa cells transiently transfected with FLAG-ZIPK were stimulated with IFN-γ or As2O3 for 12 h and stained sequentially with an anti-FLAG monoclonal antibody (top), an anti-PML polyclonal antibody (C, upper middle), or an anti-Daxx polyclonal antibody (D, upper middle) and DAPI (bottom). Antibody detection was achieved with a FITC-conjugated anti-mouse antibody (green) and a rhodamine-conjugated anti-rabbit antibody (red). Overlay results (merge) demonstrate colocalization of FLAG-ZIPK and PML or Daxx in response to IFN-γ and As2O3.