Fig 2.
PHF8 interacts with the APCcdc20 during M phase. (A and B) MCF7-pOZ-Flag-HA-PHF8 cells were used for complex purification. Tandem affinity purification (Flag followed by HA IPs) was carried out on nuclear extracts. Immunocomplexes were subjected to silver staining (A) and mass spectrometry (B). Panel B shows a summary of components of the APC complex that was identified in the PHF8 complex and their total peptide numbers. (C) The control empty vector, Flag-HA-PHF8, or Flag-HA-KIAA1718 was transfected in HEK293T cells, and lysates were subjected to HA IP and probed for CDC20. (∗, IgG heavy chain) (D) Semiendogenous IP was carried out by transiently transfecting HA-CDC20 and carrying out HA or mouse IgG (mIgG) IP with the lysates. (E and F) Endogenous IP using PHF8 (E) or CDC20 (F) antibody was carried out by using lysates from asynchronous (AS) or S- or M-phase-synchronized HeLa cells.