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Fig. 2. Import and export pathways of RXR
. (A) Effects of temperature and sodium azide on shuttling of RXR. Heterokaryons were prepared and visualized as described in Fig. 1A. Fused cells were separately incubated at 37°C or 4°C or with sodium azide (1 mM) for 2 hours. Arrows indicates the HeLa nuclei. (B) Effect of wheat germ agglutinin (WGA) on RXR import. WGA (100 µg/ml) was transfected into MGC80-3 cells by means of the Chariot protein delivery system for 2 hours and then harvested. Cell lysate, nuclear and cytoplasmic fractions were prepared as described in Materials and Methods, and were then subjected to western blotting with anti-RXR antibody. Lamin B1 and tubulin were used as loading controls. (C) Effects of leptomycin (LMB) on shuttling of RXR. NIH3T3 cells were transfected with GFP-RXR, then fused with HeLa cells. The heterokaryons were treated with or without LMB (100 µg/ml) for 2 hours. GFP-p53 was used as a positive control. The HeLa nuclei are indicated by arrows.
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