Figure 5.

Npl4 interacts with chromatin-bound Cse4 in a cdc48-3 mutant. (A) Endogenous Cse4, Npl4, and Cdc48 are enriched in chromatin of cdc48-3 strain. Chromatin was prepared from equal numbers of logarithmically growing cells. Levels of Cdc48, Npl4, and Cse4 in chromatin were monitored by Western blot analysis with anti-Cdc48, anti-Myc, and anti-HA antibodies, respectively. Histone H2B was used as marker for chromatin fraction. (B) Levels of chromatin-bound Cdc48, Npl4, and Cse4 from A were quantified in arbitrary density units after normalization to H2B. Error bars represent the standard deviation of three biological repeats. Statistical significance was assessed by unpaired t-test. (C) Npl4 interacts with chromatin-bound Cse4. In vivo interaction of Cse4 with Npl4 in chromatin was determined by Co-IP using chromatin lysates. α-Myc and α-HA antibodies were used to detect Npl4 and Cse4 on the Western blot, respectively.