Figure 1.

IFI16 is a major DDX5 interacting protein. (A) Schematic of experimental workflow. Doxycycline inducible FLAG‐DDX5 expressing HepaRG cells, grown for 48 h in the presence of doxycycline (1.0 µg/mL), followed by preparation of native whole‐cell extract (WCE) and immunoprecipitation with FLAG Ab or IgG. Immunoprecipitates were electrophoresed for 30 min using 10% SDS‐polyacrylamide gels; upon entry of protein samples into separating gel, electrophoresis was terminated; gel slices were excised and processed for in‐gel trypsin digestion and sample preparation for LC‐MS/MS. (B) FLAG‐DDX5 interacting proteins identified by LC‐MS/MS analysis. (C) CORUM [31] prediction of significant complexes from (B). (D) Immunoblots of WCE from Dox‐inducible FLAG‐DDX5 expressing cells, grown without (−) or with (+) doxycycline (1.0 µg/mL) for 48 h. WCE immunoprecipitated with IgG or FLAG Ab, and immunoblotted with indicated antibodies. A representative experiment is shown from three independent experiments (n = 3) * indicates a contaminant detected with IgG. (E) HepaRG WCE native lysates immunoprecipitated with DDX5 Ab or IgG, followed by immunoblots with indicated antibodies (n = 2).