Journal of General Virology

Fig. 4. Overexpression of MxA in HuH-7 cells does not inhibit HCV replicons but blocks THOV replication. (A)–(B) HuH-7 cells containing the HCV replicons I₃₇₇/NS3-3´ (clone 9-13) (A) or I₃₇₇/NS2-3´ (clone 11-7) (B) were transfected with an expression vector encoding MxA, seeded onto glass coverslips 24 h post-transfection, fixed 72 h later and double-immunostained for MxA (upper panels) and NS5A (lower panels) by using a rabbit polyclonal antibody directed against MxA and MAb 11H, respectively. Bars, 50 µm. (C) Parental HuH-7 cells were transfected with an expression vector encoding MxA, seeded onto glass coverslips 24 h post-transfection and subsequently infected with THOV at an m.o.i. of about 25 p.f.u./cell. The cells were fixed 9 h post-infection and immunostained for MxA (upper panels) and THOV proteins (lower panels) by using MAb 2C12 and the hyperimmune serum gp457, respectively. Bar, 50 µm. (D)–(F) Quantitative analysis of the respective experiments described in parts (A)–(C), including additional control experiments with MxA(T103A). The percentage of cells expressing both virus proteins and Mx proteins is given in relation to the total number of Mx-expressing cells. Columns and error bars represent mean values of at least three independent experiments and 95 % confidence intervals, respectively. The number of cells expressing virus proteins and Mx proteins/total number of cells expressing Mx is indicated above each column. NS5A was detected in about 90 % of non-transfected cells of clone 9-13 and in about 80 % of non-transfected cells of clone 11-7 (indicated as dotted lines); THOV proteins were found in about 80 % of non-transfected HuH-7 cells (indicated as dotted line).

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