Extended Data Figure 2: The ectopic expression of SEC14L2 confers HCV permissiveness to human hepatoma cells.

a, b, SEC14L2 is not expressed in most cell lines. a, SEC14L2 mRNA levels were measured in the indicated cells by qPCR, and the values were normalized to those of the housekeeping gene, RPS11. Shown is the fold difference from fetal hepatocytes. The results are plotted as mean ± s.e.m. of two different cell stocks. b, SEC14L2 protein expression in the indicated cells was determined by immunoblotting with SEC14L2 rabbit polyclonal antibody. β-actin is included as a loading control. c, SEC14L2-expressing Huh-7.5 cells were transfected with the indicated wild-type subgenomic replicons. After 4 weeks of selection, HCV RNA was sequenced in six individual colonies from each of the replicons. HCV RNA levels in each of these colonies were determined by qRT–PCR. d, Hep3B/miR-122 and Huh-7 cells transduced with SEC14L2 or empty vector were transfected with in vitro-transcribed RNA from the indicated wild-type replicons and selected for 3 weeks with G418 (500 µg ml⁻¹). The resulting cell colonies were stained with crystal violet. S52 GNN with a catalytically inactive mutation in NS5B polymerase was included as a negative control.