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. 1997 Nov 17;139(4):841–849. doi: 10.1083/jcb.139.4.841

Figure 5.

Figure 5

Figure 5

Unassisted nuclear migration of β-subunit occurs in the absence of exogenous addition of soluble factors in digitonin-permeabilized cell-free transport assay. (A) Digitonin-permeabilized MDBK cells were incubated with 10 μl of testing solution containing 3 pmol GFP–β-subunit (a and b) or GFP-D450-876 (c and d) in the presence (a and c) or absence (b and d) of 3 pmol GST– α-subunit and T-APC (0.1 mg/ml) for 20 min. (B) Digitonin-permeabilized MDBK cells were incubated with 10 μl testing solution containing 4 pmol of GFP–β-subunit (a–c), GFP-D450-876 (d–f), or FITC-D3-145 (g–i) in the absence (a, d, and g) or presence of either 20 pmol GTP-Ran (b, e, and h) or GDP-Ran (c, f, and i) for 15 min. After incubation, localization of β-subunit and its mutant proteins was examined after fixation of the permeabilized cells as in Fig. 3 D.