Fig. 3: COMMD4 interacts with H2B and RNF40.

a Empty vector (FLAG) and COMMD4-FLAG were immunoprecipitated from HeLa cells and the co-eluting proteins were immunoblotted with antibodies shown. b Immunoprecipitation as a, but this time cells were treated with 6 Gy IR. c A direct interaction for COMMD4 and H2B ± recombinant ATM. d A direct interaction between COMMD4 and H2B ± recombinant ATM and the ATM inhibitor (ATMi) KU-55933. e FLAG and FLAG-tagged H2B WT, and S14E mutant were immunoprecipitated from HeLa cells and the co-eluting proteins were immunoblotted with antibodies against COMMD4 and H2B. IgG heavy chain shows the loading. f Direct interaction between COMMD4 and H2B demonstrating the specific binding region within H2B. No binding was seen in the negative control with only the beads and recombinant COMMD4 (beads), while expression of His-tagged COMMD4 (COMMD4, right most lane) is seen. Schematic on the right shows peptide regions within H2B and Supplementary Table 1 shows the peptide sequences. g Direct interaction between RNF40 and H2B demonstrating the specific binding regions within H2B. Expression of recombinant RNF40 is seen in the control lane. h A three dimensional structure (PDB code 2RVQ) of the heterodimer H2A-H2B wherein H2A and H2B are shown in blue and red ribbon representation, respectively. Tails of histones are disordered and in extended conformation. The phosphorylation sites are shown as spheres.