Figure 5.

HF domains in NC2 subunits are essential for its integrity and ability to bind at cat-3 locus. (A, B) Mapping of the NC2α region responsible for the interaction with NC2β in Nc2α^KO strain with ectopically expressing wild-type Myc-NC2α or its various deletion mutants. Immunoprecipitation assays with anti-Myc antibody (A) or anti-NC2β antibody (B), the eluates were detected by western blot analysis using anti-Myc (α-Myc) and anti-NC2β (α-NC2β) antibodies. The red arrows denote specific bands, and the black arrow shows heavy chain. (C, D) Mapping of the NC2β region responsible for the interaction with NC2α in Nc2β^KO strain with ectopically expressing Myc-NC2β or its various deletion mutants. Immunoprecipitation assays with anti-Myc antibody (C) or anti-NC2α antibody (D), the eluates were detected by western blot analysis using anti-Myc (α-Myc) and anti-NC2α (α-NC2α) antibodies. The blue arrow denotes heavy chain of IgG, the black arrow shows light chain. (E, F) ChIP assays showing the binding levels of NC2α (E) and NC2β (F) at cat-3 locus in different deletion strains across NC2α coding region at endogenous locus. (G, H) ChIP assays showing the binding levels of NC2α (G) and NC2β (H) at cat-3 locus in different deletion strains across NC2β coding region at endogenous locus. Error bars show s.d. (n = 3). Significance was assessed by using a two-tailed t-test. *P < 0.05, **P < 0.01 and ***P < 0.001 vs. WT.