FIG 5.

SG proteins interact with the 3′ UTR of EV-D68 RNA to inhibit viral replication. TIA1 (A), G3BP1 (B), HUR (C), and the minireplicon were cotransfected into 293T cells, and 293T cells were cotransfected with empty vector and the minireplicon as a control. Coimmunoprecipitation was performed 48 h after transfection, and minireplicon mRNA levels were detected by qPCR before and after coimmunoprecipitation. Western blotting was performed to detect the expression of TIA1, G3BP1, and HUR. (D, E, and F) SG proteins and minireplicon deletion mutations were cotransfected into 293T cells and coimmunoprecipitated by the above method. RD cells were transfected with minireplicon (G) and mutants (H) and infected with EV-D68 24 h posttransfection, and the cells were then harvested at 0, 4, 8, 12, and 24 h postinfection. EV-D68 RNA was extracted, and qPCR was used to detect viral mRNA in the cells.