FIG 3.

Antibunyaviral activities of candidate ISGs against a panel of divergent bunyaviruses. (A) ISG expression in 10 Vero-ISG-expressing cell lines in addition to “empty vector” and naive Vero cell controls was assessed using Western blotting (assays were not established for NOS2A and RHOB). (B) For the ISGs in panel A, lentiviral vector-encoded TagRFP expression was monitored by flow cytometry. (C) Infectious virus yields from 10 Vero-ISG-expressing cell lines. Cells were infected with 15 wild-type bunyaviruses representing four families. Supernatants were collected, and virus yields were determined on Vero or BHK-21 cells. The MOIs and time points of supernatant harvest were as follows: for BUNV, CVV, KRIV, SBV, DUGV, RVFV, and SFTSV, MOI of 0.001 and 72 hpi; for OROV, MOI of 0.001 and 48 hpi; for ANAV, BMAV, BORV, CAPV, and TCMV, MOI of 0.01 and 48 hpi; for PUUV, MOI of 0.1 and 48 hpi; and for HRTV, MOI of 0.01 and 72 hpi. EMPTY, Vero cells transduced with empty SCRPSY lentiviral stocks. (D) Heat map analysis (generated by GraphPad Prism 7) of the effects of 10 ISGs on the yields of 15 bunyaviruses. (E) Summary of human ISG20's antiviral effects on 15 bunyaviruses.