Hepatitis B virus Core protein nuclear interactome identifies SRSF10 as a host RNA-binding protein restricting HBV RNA production
Fig 3
(A) Relative abundances of the 11”founder” RBPs identified in HBc nuclear complexes submitted or not to Benzonase treatment. The relative abundances of HBc binding partners have been evaluated using the iBAQ metrics [104]. For each replicate, each iBAQ value was normalized by the summed values of the 11 proteins. Error bars represent +/- SD. (B) and (C) Western blot validations in Benzonase treated and streptactin-purified and extracts. Two major isoforms of SRSF10 are visible: the upper at 37KDa and the lower at 20–22 KDa. The band indicated with an asterisk likely corresponds to a band generated by proteolytic cleavage. (D) HBc was immune-precipitated from nuclear extracts purified from liver sections from HBV-infected HuHep mice, using two different anti-HBc antibodies. Eluted proteins were analyzed by western blot using anti HBc and anti-SRSF10 antibodies. The asterisk indicates the positions of IgG heavy chain. (E) Proteins included in the gel band between 35 and 25 KDa were analyzed by MS. The table indicates the list of proteins recovered with the anti-HBc antibody that were also previously found after HBc purification on StrepTactin columns (see Fig 1D). * In the case of MLF, 1 peptide was found in the anti-IgG control IP. The other proteins were found exclusively in the anti-HBc IP.