Skip to main content
. 2010 Jul 15;21(14):2483–2499. doi: 10.1091/mbc.E10-03-0176

Figure 5.

Figure 5.

Scyl1 associates with the NPC by interacting directly with Nup98. (A) Nup98 copurifies with Scyl1. Total cell lysates prepared from HeLa cells (lane 1) were incubated with Sepharose 4B (lane 2) or α-Scyl1 coupled to Sepharose 4B (lane 3). The washed resins were boiled in sample buffer, and the eluates were subjected to Western blot analysis to detect Nup98 (first row), Nup107 (second row), and Scyl1 (third row). The proteins were visualized directly by SYPRO Ruby staining of the blot (fourth row). (B) Scyl1 interacts directly with Nup98 in vitro. Bound GST-Nup98 (20 μg; 161 pmol) (lane 3) or GST (lane 4) was incubated with a twofold molar excess of Scyl1 (28 μg; 322 pmol). The washed resins were boiled in sample buffer, and the eluates and purified Scyl1 (lane 1) and GST-Nup98 (lane 2) were subjected to SDS-PAGE. Scyl1 (first row) and GST-Nup98 (second row) were detected by Western blot analysis followed by Coomassie Blue staining of the blot (third row).