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. 2018 Oct 19;8:15486. doi: 10.1038/s41598-018-33861-6

Figure 3.

Figure 3

SLC3A2 expression is upregulated by NS3/4A-induced oxidative stress. (A) Huh7.5 cells were transiently transfected with either vector (−) or increasing amounts of Myc-tagged core, NS3, NS3/4A, NS4B, NS5A, NS5B, or V5-tagged E2, E1E2 expression plasmid, respectively. At 36 h after transfection, total cell lysates were immunoblotted with the indicated antibodies. (B) Huh7.5 cells were infected with either VSVpp harboring HCV envelope protein or HCVpp harboring HCV envelope protein derived from H77 or JFH1 for 6 h. At 48 h postinfection, SLC3A2 protein expression levels were analyzed by immunoblot analysis. (C) (Upper panel) Huh7.5 cells were transiently transfected with empty vector, Myc-tagged core, NS3, NS4B, NS5A, NS5B, or V5-tagged E2, E1E2 expression plasmid, respectively. At 36 h after transfection, mRNA levels of SLC3A2 were determined by qRT-PCR analysis. (Lower panel) Huh7.5 cells were transiently transfected with either empty vector or increasing amounts of NS3/4A expression plasmid. At 36 h after transfection, mRNA levels of SLC3A2 were determined by qRT-PCR analysis. (D) Huh7.5 cells were transfected with either vector or NS3/4A expression plasmid. At 36 h after transfection, cells were treated with increasing amount of NAC. At 24 h after treatment, both protein (left panel) and mRNA levels (right panel) were analyzed by either immunoblot assay or qRT-PCR, respectively.