Skip to main content
. 2012 May 24;8(5):e1002696. doi: 10.1371/journal.ppat.1002696

Figure 3. Infectivity titers and replication capacity of HCV NS5A LCSII mutants.

Figure 3

(A) Infectivity titers in supernatant after transfection of Huh7.5 cells with original NS5A genotype recombinants or LCSII mutants. For the residue 343 mutants, J4 and JFH1 recombinants are shown as representative examples, since no significant effect on titers was observed for any of the NS5A genotype 1–7 recombinants. For comparison, JFH1(2a) was included in each experiment. The lower limit of detection in the experiments shown was 102.4 FFU/mL (indicated by the y-axis break). (B) As a measure for HCV RNA replication, S29 cells were transfected for 4 hours with original NS5A genotype recombinants and P346A/P351A/P354A mutants, and intracellular HCV Core levels were measured after 4, 24, 48 and 72-hours. Values were normalized for transfection efficiency using 4-hour Core amounts of the respective recombinant. The JFH1(2a) NS5BGND mutant was included as a replication negative control. (C) Intra- and extracellular infectivity titers 48 hours after transfection of S29 cells with original NS5A genotype recombinants and P346A/P351A/P354A mutants. Intracellular titers are given per well of 4×105 transfected S29 cells. #: no FFU were detected. Colored boxes indicate the NS5A isolate and (genotype). Error bars indicate SEM of triplicate determinations, except for in (B) where JFH1(2a) was included in duplicates.