Fig. 1: Proteome analysis of HCV-induced DMVs identifies AGPATs as host dependency factors critically contributing to viral replication.

a Experimental approach used to purify DMVs from HCV-replicating cells. b Volcano plot of differentially enriched interactors of NS4B and calnexin (CNX). Q-values were calculated using the limma software package and corrected for multiple hypothesis testing. Viral proteins are highlighted with red letters. A magnified view with protein hits labeled is given in Supplementary Fig. 1B. c A total of 139 genes were selected from the DMV proteome and validated by siRNA screening (3 siRNAs per gene). CD81, PI4KA and Rluc were used as positive controls; NC (negative control)1, NC2, GFP and mock infection served as negative controls. A summary of the screening is given in Supplementary Data 2. d Endogenous AGPAT1 and 2, but not AGPAT3 are contained in NS4B-associated membranes. Membranes were purified from naive Huh7-Lunet cells (-), or Huh7-Lunet cells containing a subgenomic replicon without or with an HA-tag in NS4B (NS4B-wt and NS4B-HA, respectively). Captured proteins were analyzed by western blot, along with the input (2%). α-tubulin served as loading control. Two biologically independent experiments showed similar results. e Colocalization of NS4B with AGPAT1 and 2. Huh7-Lunet cells stably expressing AGPAT1- or AGPAT2-EGFP were transfected with in vitro transcripts of the HCV genome Jc1 and fixed 48 h post-transfection. Two biologically independent experiments showed similar results. Source data for panels c and d are provided as Source Data file.