Examination of TNF-α-induced NF-κB p65 nuclear translocation and degradation of IκBα in HTNV-infected cells. A549 cells were mock infected (M) or infected with HTNV (V) at an MOI of 5. On day 5, mock- and HTNV-infected cells were left untreated (−) or treated (+) with 50 ng/ml of TNF-α for 15 min. (A) After fixation, cells were stained with antibodies against NF-κB p65 (green) and HTNV N protein (red) and stained with DAPI to highlight nuclei (blue). (B) Cell lysates were separated into cytoplasmic and nuclear fractions. (C) Total cell lysates were prepared for immunoblotting, and proteins were transferred onto PVDF membranes. Blots were probed with antibodies against IκBα, p50, p65, HTNV N protein, GAPDH, or histone H2B.