FIG 7.
Effect of ectopic expression and downregulation of expression of selected host proteins on infectious virus yield. HEK293T cells in duplicate wells in a 24-well plate were transfected with the Myc-tagged hnRNP D45 isoform, hnRNP C1, hnRNP E1, hnRNP I, hnRNP L, NH-HuR, and ECFP-G3BP1 plasmid DNA constructs (1 μg/well) or the corresponding siRNAs (25 pmol/well) for 48 h, followed by infection with 400 FFU of the RRV strain of rotavirus. Nontarget fluorescent Accell siRNA was used as a control. At 8 hpi, cell lysates were prepared in DMEM by the freeze-thaw method. MA104 cells were infected with serially diluted lysates from the transfected-infected HEK293T cells, and the progeny virus titers in two wells each from two independent experiments were determined by an ELIFA as described in Materials and Methods. MAb 631/9 against VP6 was used for detection, and deep-brown foci were counted under a microscope.