Fig 9. CRM1-mediated export complexes accumulate at ALPCs when the disassembly of these export complexes is inhibited by transient expression of RanQ69L mutant.

(A and B) HeLa cells were transfected with the construct encoding FLAG-tagged Ran wild-type (WT) or RanQ69L GTPase mutant (Mut), double-labeled with anti-FLAG and RanGAP1 antibodies (A) or mAb414 (B) and analyzed by immunofluorescence microscopy. (C-E) HeLa cells were co-transfected with the constructs encoding Rev-GR-GFP and FLAG-tagged Ran WT or RanQ69L. The transfected cells were incubated with 1 μM dexamethasone to induce the nuclear accumulation of Rev-GR-GFP, washed with fresh medium to remove dexamethasone to initiate nuclear export, stained with anti-RanGAP1 antibodies (C), anti-FLAG antibodies (D), or both anti-RanGAP1 and anti-FLAG antibodies (E), and analyzed by immunofluorescence microscopy. Bar, 10 μm. The boxes at the top-left corner of each image reveal the enlarged version of inlets.