Gs 9620

Editing was performed as described above in activated primary CD4+ T cells with the following exceptions: crRNA guides used were NTC-02, MARCH5-01, TRAF2-01, TRAF3-03, UHRF1-01, and ZFP91-02 (designed by Horizon, Table S2), no stimulation beads were used, and complete media were RPMI-1640 media (Corning) with 10% FBS (Gibco), 50 μg/mL penicillin-streptomycin (Fisher), and 2 mM L-Glutamine (Corning). Then, 300,000 JLat cells were resuspended in 20 µL Lonza electroporation buffer SE (Lonza) and electroporated with code CM-137. Cells were fed with complete media every 2–3 days until drug treatment. Eight days after electroporation with crRNPs, JLat cells were treated with 10 ng/mL TNFalpha (PeproTech), 100 nM AZD5582 (a generous gift from Sumit Chanda’s lab), 100 nM GS-9620 (Cayman Chemical), 10 µg/mL PHA (a generous gift from Melanie Ott’s lab), 16 mM PMA (Ott lab) + 0.5 µM Ionomycin (Ott lab), and 625 nM JQ1 (Ott lab) or untreated with complete media. Cells were collected 24 hours after treatment by resuspension in 1% paraformaldehyde in PBS.

Neutralizing antibodies against mouse IFN- alpha receptor or IFN- gamma (Bio X Cell) were i.p. injected at a dose of 150 μg/mouse into 1-week-old wt-129 mice, either before or after i.p. inoculation with EV-A71 clinical isolate F23 (10⁸ PFU/mouse). The neutralizing antibody against the IFN-alpha receptor was injected 1 day before EV-A71 inoculation, followed by repetitive antibody treatments on 1 and 3 dpi. Recombinant mouse IL28A and IL28B were both purchased from R & D Systems®. Various doses of mouse IL28 was i.p. injected into mice at dpi = 1 (Additional file 4: Figure S4a). Recombinant mouse IFNαA (PBL Assay Science Co.), GS9620 (Cayman Chemical) and R837 (InvivoGen), were orally or i.p. injected into wt-129 mice infected with EV-A71, at the dose of 300 IU/g (mouse IFNαA), 3.0 μg/g (GS-9620), 4.5μg/g (GS-9620), and 5.0 μg/g (R837), respectively.