Fig. 3

ATF-2 and NF-κB are activated in poliovirus-infected cells. a. Phosphorylation of ATF-2. Whole cell lysates prepared from uninfected or poliovirus-infected cells were analyzed by immunoblotting. Phosphorylated ATF-2 (phospho-ATF-2) was detected by using an antibody specific for the phosphorylated form of ATF-2. The membrane was also probed with an antibody to β-actin to show equivalent loading of protein lysates. b. Subcellular localization of NF-κB. Cells that were mock-infected or had been infected with poliovirus for the indicated amount of time were analyzed by immunofluorescence with antibodies to detect the p65 subunit of NF-κB. Panels labeled NF-κB show cells stained with a rabbit polyclonal antibody to detect the p65 subunit of NF-κB using TRITC filter, while the panels labeled DNA show the same fields stained with Hoechst to reveal the nuclei. c. Steady state levels of NF-κB. Whole cell lysates prepared from uninfected or poliovirus-infected cells at an MOI 50 (lanes 2–4) or 100 (lanes 5–7) were analyzed by immunoblotting. The p65 subunit of NF-κB was detected by probing the membrane using antibody specific for p65. The membrane was also probed with an antibody to β-actin to show equivalent loading of protein lysates. Arrows indicate cleavage products produced following infection. hpi: hours post infection