RNA helicase A is necessary for robust REV-A PCE activity and HTLV-1 Gag production

<p><b>Copyright information:</b></p><p>Taken from "RNA helicase A interacts with divergent lymphotropic retroviruses and promotes translation of human T-cell leukemia virus type 1"</p><p></p><p>Nucleic Acids Research 2007;35(8):2629-2642.</p><p>Published online 10 Apr 2007</p><p>PMCID:PMC1885656.</p><p>© 2007 The Author(s)</p> COS cells were transfected with siRNAs targeting RHA or with scrambled sequences (Sc), incubated for 72 h and co-transfected with the siRNAs and indicated PCE reporter plasmid or HTLV-1 provirus. After 48 h, total cellular protein was harvested and subjected to immunoblot with antiserum to RHA to assess downregulation, Gapdh antiserum to control for sample loading, and F-luc to control for non-specific effects of siRNAs. () Measurements of PCE activity by Gag ELISA on the total cellular protein are summarized below immunoblot. The detection limit of the assay was (25 pg/ml). Results are representative of three independent experiments. () Lysates from three independent transfection assays were used for immunoblot for RHA and Gapdh. Results of HTLV Gag ELISA on cell-associated or cell-free protein preparations are summarized below immunoblot