De novo ferritin synthesis in poliovirus-infected cells. (A) HT1080 cells were infected with the Sabin-like type I strain of poliovirus at a multiplicity of infection (MOI) of 10. At the indicated time intervals post-infection (h), cell lysates were prepared and analyzed by Western blotting with eIF4GI (top) and phospho-eIF2α (bottom) antibodies. (B) HT1080 cells were infected with the poliovirus (MOI of 10) for 15 h, treated with hemin for 6 h, and metabolically labeled with [35S]methionine/cysteine for 2 h. Cell extracts (1000 μg) were subjected to immunoprecipitation with 30 μg of ferritin antibody. Immunoprecipitated material and the IP supernatants were analyzed by SDS-PAGE on a 12% gel. The ferritin bands and the lanes corresponding to host and viral protein synthesis were visualized by autoradiography. (C) Analysis of H-ferritin mRNA from whole-cell lysates by qPCR.
