Fig. 2.

Distribution of the subunits and reconstitution of the Wave-2 complex. (A) A HeLa extract was loaded on a sucrose gradient, and the fractions were immunoblotted with antibodies targeting the different subunits. The complex peaks at ≈11 Swedbergs (fractions 13 and 14) where all five proteins could be detected. (B) Reconstitution of the complex by in vitro translation. The genes encoding the five subunits were cotranslated in vitro in the presence of [³⁵S]methionine, and the mixture was separated on a sucrose gradient. Nap is the limiting component in the mixture and is highly enriched at the size of the native complex, 11 S. The other subunits also were incorporated in the reconstituted complex at 11 S.