Effect of Leptomycin B (LMB) treatment on RanBPM nuclear export.

<p><b>A)</b> RanBPM shRNA Hela cells transfected with 840 WT NLS1/NES were treated with ethanol (EtOH) or 20nM LMB, fixed following 3h of treatment and stained with DAPI. Subcellular localization was scored as either N>>C (completely nuclear), N>C (nuclear greater than cytoplasmic, N = C (nuclear equal to cytoplasmic), C>N (cytoplasmic greater than nuclear, or C>>N (completely cytoplasmic). Data represent averages from three separate experiments, each assessing approximately 50 cells. Error bars represent SD. ***, <i>P</i><0.001; **, <i>P</i><0.01; *, <i>P</i><0.05. <b>B)</b> Hela cells were treated with EtOH or 20nM LMB and incubated for 16h. Cells were fixed and processed for immunostaining with antibodies to RanBPM and cyclin B1 and nuclei stained with DAPI. At least 100 cells were scored as N>C (nuclear greater than cytoplasmic), N = C (nuclear equal to cytoplasmic), or C>N (cytoplasmic greater than nuclear). Data represent averages from three separate experiments, each assessing approximately 100 cells. Error bars represent SD. Scale bar: 10μm.