Figure 3. EM analysis of the Nup84... | Rockefeller University Press

Figure 3.

$Figure 3. EM analysis of the Nup84 complex. (A–C) Affinity-purified Nup84 complexes corresponding to the full-length complex (A), Nup85(233–744) truncation (B), and the Nup84 complex lacking Nup133 (C) were purified by either sucrose gradient (A and B) or size-exclusion chromatography (C). Coomassie-stained SDS-PAGE of the precipitated input (In) and collected fractions are shown. Fractions indicated with arrows were analyzed by negative-stain EM. Class averages for each complex are shown in the bottom part of each panel. The missing mass observed in Nup85(233–744) truncation (B) and the six-member complex (C) are indicated with arrowheads. WT, wild type. Bars, 10 nm.$

EM analysis of the Nup84 complex. (A–C) Affinity-purified Nup84 complexes corresponding to the full-length complex (A), Nup85(233–744) truncation (B), and the Nup84 complex lacking Nup133 (C) were purified by either sucrose gradient (A and B) or size-exclusion chromatography (C). Coomassie-stained SDS-PAGE of the precipitated input (In) and collected fractions are shown. Fractions indicated with arrows were analyzed by negative-stain EM. Class averages for each complex are shown in the bottom part of each panel. The missing mass observed in Nup85(233–744) truncation (B) and the six-member complex (C) are indicated with arrowheads. WT, wild type. Bars, 10 nm.