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. 2016 Jul 11;90(15):6832–6845. doi: 10.1128/JVI.00352-16

FIG 5.

FIG 5

The C-terminal domain of ISG12a domain II is critical for NS5A degradation. (A) FL-neo cells were transfected with different dose of p3×FLAG-ISG12a-DII plasmid for 48 h. NS5A was detected by Western blotting. (B) FL-neo cells were transfected with p3×FLAG-ISG12a-DII plasmid for 36 h, followed by treatment with MG132 (25 μM) for 6 h. NS5A was detected by Western blotting. (C) FL-neo cells were transfected with the indicated plasmids for 48 h. NS5A protein was detected by Western blotting. (D and E) Huh7.5 cells were infected with HCV for 6 h and then transfected with the indicated plasmids for 48 h. Intracellular HCV RNA (D) and extracellular infectious virus particles (E) were detected by real-time PCR and FFU assay, respectively. Error bars represent SD from triplicate experiments. *, P < 0.05. (F) Schematic representation of the truncations of ISG12a-DII-C. (G to I) Western blotting of lysates from HEK293T cells expressing exogenous NS5A and ISG12a domains (G) and FL-neo cells expressing exogenous ISG12a domains (H and I) for 48 h. Flag was the tag used.