FIG. 3.

Rapid nuclear translocation of NF-κB in astrocytes after TMEV infection. Nuclear translocation of NF-κB is apparent as early as 15 min after TMEV infection. TNF-α was used as a positive control. (A) NF-κB translocation was determined by immunohistochemistry using anti-p65 antibody for NF-κB and DAPI counterstaining for nuclei locations at 15 min postinfection with an MOI of 10. (B) Viral infectivity was assessed by in situ hybridization of TMEV-infected astrocytes (6 h) with digoxigenin-labeled antisense VP4 probe followed by development with alkaline-phosphatase-labeled antidigoxigenin. Arrowheads indicate the virus-infected cells. (C) Specific-binding ability of nuclear NF-κB was determined by EMSA at 30 min postinfection with an MOI of 10. Astrocytes were treated with TNF-α (10 ng/ml) for 30 min and used as a positive control. CON, control; TV, TMEV-infected; TNF, TNF-α treated.