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. 2018 Jun;519:33–41. doi: 10.1016/j.virol.2018.03.026

Fig. 1.

Fig. 1

Construction and analysis of the expression of E1E2-Flag glycoproteins. A. Schematic diagram of E1E2-Flag. The numbering corresponds to the amino acid positions in the polyprotein of the genotype 1a H77 strain. TMD – trans-membrane domain; HVR-1 – hypervariable region 1. B. Western blot detection of E2 wt and E2–Flag protein expression in 293T cells 24 h after transfection. Detection using anti-E2 AP33 (left column) or anti-Flag M2 (right column) mAbs, followed by anti-mouse IgG AP conjugate and NBT/BCIP substrates. Mock-transfected 293T lysates were used as a negative control. Loaded amounts were normalized for total protein content. C. Detection of gt1a E2 wt and E2-Flag proteins by immunohistochemical staining in fixed 293T cells. 48 hrs post transfection with either E1E2 wt or E1E2–Flag expression plasmids, the cells were fixed and probed with anti-E2 AP33 or anti-Flag M2 mAbs. Mock transfected HEK cells were used as a negative control.