Fig 3.

Prevention of BMRF1-dependent nuclear transport of BALF5 DNA polymerase catalytic subunit by the Hsp90 inhibitor radicicol. HeLa cells were transfected with the indicated combinations of pcDNA-FLAG-BALF5, pcDNA-BMRF1, and pcDNA-HA-Hsp90 in the absence or presence of 1 μM radicicol. After fixation 24 h posttransfection, the cells were immunostained with anti-FLAG (green) and anti-HA (yellow) antibodies (A), with anti-BMRF1 (red) and anti-HA (yellow) antibodies (B), and with anti-FLAG (green), anti-BMRF1 (red), and anti-HA (yellow) antibodies(C). Nuclei were stained with DAPI (blue). (D) Numerical analysis of the localization pattern of BALF5 in transfected cells. The proportions of cells showing each localization pattern are expressed as percentages out of the examined cells (more than 100). The images in the upper panels represent the typical BALF5 localization patterns. (E) Effect of Hsp90 inhibitor. HeLa cells were treated with 0.5 μM radicicol for 24 h and harvested. Clarified cell lysates were prepared, separated by SDS–10% PAGE, and applied for Western blot analyses with anti-Chk1 protein polyclonal antibody.