FIGURE 2.

EV-A71 2B reduced levels of KPNA1 and p-STAT1/KPNA1 complex. (A) Reduction of KPNA1 levels by EV-A71 viral proteins. HeLa cells were infected with EV-A71 at an MOI of 1 or transfected with plasmids (2.0 μg) expressing HA-tagged viral proteins. Cell lysates were prepared for western blot analyses with antibodies for HA tag or KPNA1. (B) Reduction of KPNA1 levels by viral 2A, 2B, and 3C. HeLa cells were infected with EV-A71 or transfected with plasmids expressing HA-tagged 2A, 2B, or 3C. Cell lysates were prepared for western blot analyses with antibodies for HA tag and KPNA1. (C) Reduction in levels of the p-STAT1 and KPNA1 complex in IFN-α-stimulated cells by 2B. HeLa cells were transfected with plasmids expressing HA-tagged 2B (2.0 μg). At 36 h post transfection, the cells were treated with IFN-α (25 ng/μL) for 2 h. Cell lysates were prepared for co-immunoprecipitation (Co-IP) with antibodies for HA, p-STAT1, or KPNA1. Normal isotype IgG was used as negative control. Subsequent western blot analyses were performed with antibodies for HA, p-STAT1, or KPNA1. The levels of inputs in the whole cell lysates (WCL) were also shown together with β-Tubulin. (D) EV-A71 2B reduced the levels of the exogenous p-STAT1 and KPNA1 complex. HeLa cells were transfected with plasmids expressing FLAG-KPNA1 (2.0 μg), EGFP-STAT1 (2.0 μg), and HA-2B (2.0 μg). At 36 h post transfection, the cells were treated with IFN-α (25 ng/μL) for 2 h. Cell lysates were prepared for Co-IP with an anti-FLAG antibody and subsequent western blot analyses with antibodies for EGFP tag. The levels of inputs in WCL were also shown with antibodies specific for FLAG, EGFP, or HA tags.