Fig. 1
HIRA homooligomerizes in cells and forms a homotrimer in vitro. a YFP constructs of human HIRA and mutants. The WD40 repeat (aa 1–369), B (aa 439–475) and C (aa 763–963) domains involved in UBN, ASF1a, and CABIN1 interactions, respectively, are shown. Star indicates single amino acid substitution with alanine. The co-immunoprecipitation efficiency of endogenous HIRA or HIRA-HA (HIRA coIP) is 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 YFP-tagged and HIRA-HA proteins. d Western blot analysis of anti-GFP-immunoprecipitates from U2OS nuclear extracts expressing YFP-tagged proteins. In b, c and d, input corresponds to 10% of nuclear extract used for each experiment. e Equilibrium sedimentation of recombinant proteins HIRA(661–1017), CABIN1 full length (FL) and HIRA(661–1017) + CABIN1 FL. Theoretical (open symbols) and experimental (closed symbol) curves are shown
