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. 2015 May 20;62(2):397–408. doi: 10.1002/hep.27847

Figure 5.

Figure 5

Low level of viral competition argues against a strong overlap of host factors between HAV and HCV. (A) Schematic representation of subgenomic reporter replicon constructs used for coelectroporation. The R‐LucJFH X‐tail Con1 chimera is characterized by reduced replication efficiency. (B‐D) Coelectroporation of HAV and HCV points to a low level of viral replicase competition. Huh7‐Lunet cells were either electroporated with single‐reporter replicons or coelectroporated with different combinations of HAV and HCV subgenomic reporter replicons (CoEpo). To distinguish between different coelectroporated constructs, firefly and renilla luciferase reporter replicons were combined. Then, 4, 24, 48, and 72 hours upon electroporation, firefly and renilla luciferase activity were determined, normalized to the 4‐hour value, and depicted in logarithmic scale (B: firefly luciferase activity; C and D: renilla luciferase activity). Graph depicts mean and standard deviation of a representative experiment (n = 2). (B) LucHAV, (C) R‐LucJFH, (D) R‐LucJFH X‐tail Con1, (E) IF analysis of Huh7‐Lunet cells 72 hours upon coelectroporation with HAV and HCV replicon constructs (upper two panels) or mock electroporation (lower panel) using mouse monoclonal anti‐HCV‐NS5A antibodies and a rabbit polyclonal anti‐HAV‐3C serum. Right panel represents a magnification of the boxed areas. Scale bar, 10 µm. Abbreviations: F‐Luc/Luc, firefly luciferase; R‐Luc, renilla luciferase.