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. 1997 May 19;137(4):797–811. doi: 10.1083/jcb.137.4.797

Figure 3.

Figure 3

The NES region of Kap95p mediates nuclear export. (A) Alignment of a region of Kap95p with the NES sequences of HIV-1 Rev, PKI, TFIIIA, and Gle1p is shown. (B) GST-Kap95-(1-77) and GST-Kap95-(1-77)ΔNES fusion proteins were purified and conjugated to FITC. The fluorescent conjugates were individually coinjected into the nuclei of COS-1 cells with Texas red–HSA. After a 45min incubation at either 37°C or 4°C, the cells were fixed and examined by fluorescence microscopy. The Texas red– HSA remained nuclear localized (bottom row); however, the FITC-GST-Kap95-(1-77) moved to the cytoplasm (top, left) when incubated at 37°C. The FITC-GST-Kap95-(177) was confined to the nucleus at 4°C incubation (top, middle). The fusion lacking the NES motif (FITC-GSTKap95-(1-77)ΔNES) remained nuclear even at 37°C (top, right). Bar, 10 μm.