Due to the RNA nature of their genomes, influenza viruses have to utilize many RNA-binding proteins (RBPs) of both viral and host origin, for their replication. To uncover the comprehensive vRNA-host protein interactions, we performed affinity purification coupled with mass spectrometry (AP-MS) analysis of influenza vRNA complexes. The eight vRNA segments of H7N9 were transcribed and individually labeled with biotin in vitro and incubated with IAV virus-infected THP-1 cells, and vRNA complexes were enriched streptavidin magnetic beads and analyzed by mass spectrometry