Fig. 2

UBN1 mediates the interaction between HIRA subunit and histone variant H3.3. a UBN1 mediates the specific interaction between HIRA and H3.3. Interaction between HIRA subunit and H3.3 is analyzed by LacO-LacI targeting system in A03_1 or A03_1/Flag-UBN1 cell lines. A green focus indicates the interaction between the histones with chaperones. Scale bar, 10 μm. b Interaction between HIRA subunit and H3.3 is analyzed in the presence of UBN1 or Cabin1 by Western blot analysis of anti-Flag immunoprecipitates. c Both WD repeat domain and B domain of HIRA are involved in the interaction with H3.3. Top panel, schematic presentation of full length and truncation mutants of human HIRA; bottom panel, interaction between truncation mutants of HIRA subunit and H3.3 is analyzed in A03_1/Flag-UBN1 cell line. Statistic results are shown in Additional file 1: Figure S2D. Scale bar, 10 μm. d The effect of UBN1 (aa1-213) on the interaction between HIRA (aa1-481) and H3.3 is analyzed by GST pull-down followed by Coomassie staining. Input corresponds to 10% proteins. e The interaction between HIRA subunit and H3.3 is independent of Asf1a. The effect of Asf1a on the interaction between HIRA subunit and H3.3 is analyzed by Western blot analysis of anti-Flag immunoprecipitates. * indicates Myc-UBN1; ** indicates Myc-Asf1a