FIGURE 3.

NVL2^UD is a nucleolin binding domain. A, in vitro binding assay of GST-NVL2^UD with a HeLa cell extract. GST-NVL2^UD(1–93)-associated proteins were separated by SDS-PAGE and stained with Coomassie Brilliant Blue. The band indicated by an arrow was subjected to the MS/MS analysis. B, Western blot of HeLa proteins captured by GST-NVL2^UD. GST or GST-NVL2^UD(1–74) was incubated with either HeLa whole cell extract (WCE) or buffer, and co-purified proteins were electrophoresed. The proteins were blotted onto a PVDF membrane and then detected by an anti-nucleolin antibody. The epitope for the antibody is residues 271–520 of human nucleolin. C, domain architecture of human nucleolin. The arrow marks the position of the peptide sequence detected by the MS/MS analysis. The RRM domains and the GAR domain are shown as black and gray boxes, respectively, with the domain names.