Fig. 2.

HIRA homooligomerization is required for CABIN1 interaction. a YFP constructs of human HIRA and mutants. The amino acids R227 and I461 critical for UBN and ASF1a interactions, respectively, are shown. Star indicates single amino acid substitution (R227 with K or I461 with D). The co-immunoprecipitation efficiencies of both endogenous HIRA (HIRA coIP) and CABIN1 (CABIN1 coIP) are indicated for each construct, “+” indicates that the efficiency of coIP is similar to the one obtained with the wt HIRA construct and “−” indicates that the efficiency of the coIP is decreased. b Western blot analysis of anti-GFP-immunoprecipitates from U2OS nuclear extracts expressing YFP-tagged proteins. c Western blot analysis of anti-GFP-immunoprecipitates from U2OS nuclear extracts expressing both HIRA-YFP and HIRA-HA proteins prepared from cells treated with siRNAs control or CABIN1. In b and c, input corresponds to 10% of nuclear extract used for each experiment