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D23A p10 does not inhibit RanGDP nuclear accumulation, but does inhibit BSA–NLS nuclear accumulation in vivo. HeLa cells were coinjected in the cytoplasm with the following: FITC–RanGDP (1 mg/ml), TRITC-BSA–NLS (2 mg/ml), the injection marker Cascade blue–labeled BSA (Molecular Probes) (1 mg/ml), and unlabeled wt or mutant p10 (2.2 mg/ml). The cells were incubated 5 min at room temperature after microinjection, and then fixed for observation.
