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. 2009 Aug 13;28(17):2541–2553. doi: 10.1038/emboj.2009.200

Figure 4.

Figure 4

Effects of WGA and Impβ45-462 on facilitated and passive NPC passage. Permeabilised cells were either left untreated (left row) or pre-incubated with 0.1 mg/ml wheat germ agglutinin (middle row) or with 3 μM of the dominant-negative importin β45-462 fragment (right row). Thereafter, indicated influx experiments were performed. Import of IBB-GFP was performed in the presence of importin β, Ran and a GTP-regenerating system. All other import mixtures comprised only the fluorescent proteins. WGA and Impβ45-462 inhibited both, facilitated NPC passage (of IBB-GFP·importin β or NTF2) and passive nuclear entry of e.g. GFP. The inhibitory effects were strongest towards the large (132 kDa) IBB-GFP·importin β complex, strong towards GFP-sized objects (NTF2, NTF2W7R, GFP), and non-significant towards the smallest substrate, an 11 amino acid long peptide. For quantitation, see Figure 5.