FIG 3.

Overexpression of SG proteins inhibits the replication of EV-D68. (A) HUR, TIA1, G3BP1, or an empty vector control were overexpressed in RD cells. RD cells were infected with EV-D68 (MOI of 0.1) 24 h after transfection, and cells were harvested after infection at 0, 4, 8, 12, and 24 h. RNA was extracted, and qPCR was used to detect viral mRNA in cells. (B) HUR, TIA1, G3BP1, and an empty vector control were overexpressed in RD cells. Cells were infected with EV-D68 (MOI of 0.1) 24 h after transfection. Cells were harvested 12 and 24 h after infection, and the cells were repeatedly thawed three times in order to assess the 50% tissue culture infective dose (TCID₅₀) virus titer. (C) HUR, TIA1, G3BP1, or an empty vector control were overexpressed in RD cells. Cells were infected with EV-D68 (MOI of 0.1) 24 h after transfection, and cells were harvested after 12 and 24 h postinfection for SDSPAGE gel electrophoresis. Western blotting detected the expression of viral protein VP1 in the cell lysate. (D) 293T cells were transfected with siG3BP1 (100 nM) using Lipofectamine 2000. The cells were infected with EV-D68 (MOI of 0.1) 48 h after transfection, and the cells were harvested at 0, 4, 8, 12, and 24 h after infection. (E) Western blot analysis detected the expression of viral protein VP1 in the cell lysate. Knockdown of G3BP1 was assessed by Western blotting.