Analysis of pseudorevertants selected with NS4B AH1 mutant K18A.

<p>(<b>A</b>) Selected amino acid changes identified following transfection of a selectable subgenomic replicon harboring the NS4B AH1 K18A mutation (see <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004501#ppat-1004501-t001" target="_blank">Table 1</a>) were reengineered into a subgenomic HCV replicon harboring a luciferase reporter gene. <i>In vitro</i> transcribed RNAs were electroporated into Huh-7.5 cells, followed by luciferase activity measurement at 4, 24, 48 and 72 h post-electroporation. Relative light units (RLU) were normalized to values measured at 4 h. Results from two independent experiments were pooled (n = 8 for each construct and time point). For statistical analyses, RLU measured at 48 h were compared to the 48-h values obtained with mutant K18A. *, P<0.001. (<b>B</b>) Structure models of the different AH1 sequences were established by using Swiss-PdbViewer 4.01 software <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004501#ppat.1004501-Guex1" target="_blank">[69]</a> and drawn by using VMD 1.9 software <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004501#ppat.1004501-Humphrey1" target="_blank">[68]</a>.