FIG. 2.

Affinity purification of the hPrp19/CDC5L complex. Complexes present in nuclear extract prepared from HeLa cell lines expressing FLAG/HA-tagged AD002 or SPF27 were immunoaffinity selected using anti-FLAG-M2-agarose and subsequently eluted under native conditions with FLAG peptide. Affinity-purified complexes were then fractionated on a linear 5-to-20% glycerol gradient. Proteins present in each gradient fraction after affinity purification from the FLAG/HA-AD002 cell line (A) and the FLAG/HA-SPF27 cell line (B) were analyzed by SDS-PAGE and stained with Coomassie blue. The molecular masses of marker proteins are indicated on the left. Proteins were identified by mass spectrometry and are indicated by arrows as follows: CDC5L (gi 11067747), HSP73 (gi 5729877), CTNNBL1 (gi 18644734), PRL1 (gi 4505895), hPrp19 (gi 7657381), AD002 (gi 6523797), and SPF27 (gi 5031653).