Fig. 3

S109 decreases CRM1 protein expression and inhibits CRM1-dependent nuclear export in glioma cells. a U87 and U251 cells were incubated with S109 for 12 h, and the CRM1 protein levels were analyzed by immunoblotting. b U87 cells were treated with LMB for 12 h and then evaluated by western blot analyses. c U87 cells were treated with S109 (2 μM) for 12 h. The drugs were then washed out, and fresh medium was added. The cells were further incubated for the indicated times, and whole-cell lysates were analyzed by immunoblotting. d U87 cells were treated with S109 in the presence or absence of LMB (2 nM) for 12 h. The CRM1 protein levels were examined by western blot analysis. e U87 cells were treated with LMB (2 nM) and S109 (1 μM) in the presence or absence of MG132 (10 μM) for 12 h. Total proteins were extracted and used for immunoblotting analysis. f, g U87 and U251 cells were transiently transfected with the NES-GFP plasmid and treated with LMB and S109 at the indicated concentrations for 2 h. Fixed cells were stained with DAPI and analyzed by fluorescence microscopy. h, i U87 and U251 cells were incubated in the presence or absence of S109 for 2 h. The cells were then fixed, stained with RanBP1 antibody and DAPI, and analyzed by fluorescence microscopy