FIGURE 3.
Gad8 physically interacts with the MBF complex. A, detection of physical interactions between Res1, Res2, or Cdc10 and Gad8. Protein extracts from wild-type cells expressing the indicated chromosomally tagged proteins were immunoprecipitated with anti-MYC antibody. Western blotting was performed using either anti-HA or anti-MYC to detect the presence of tagged proteins within the immune complexes. The expression levels of the indicated proteins before immunoprecipitations is shown (Input). Act1 was used as a loading control. B, the kinase activity of Gad8 is not required for its interaction with Res1, Res2, or Cdc10. Physical interactions between Res1, Res2, or Cdc10 and gad8-KD (gad8K259D; a kinase-dead allele) were detected as above. IP, immunoprecipitation.