Real time primers

RNA was isolated from TG and converted into cDNA as described (9 (link)). Relative gene expression was calculated by the standard 2^−ΔΔCt method, standardized reference genes, and normalized to WT UI controls following semi-quantitative real time PCR using a CFX Connect thermocycler (Bio-Rad, Hercules, CA). Viral transcripts were amplified using iTaq supermix (Biorad) with real-time primers from Integrated DNA Technologies (Coralville, IA). Primer sequences are listed in Supplemental Table I. PrimePCR technology (Bio-Rad) was utilized for ISG transcript expression studies according to the manufacturer’s instructions. Profiles of transcript expression represented by the cluster image map (or heat map) data supplement were generated using the National Cancer Institute’s CIMminer tool freely accessible online.

RNA was extracted by using TRIzol Reagent (Invitrogen). Quantitative RT-PCR was performed with PrimeScript 1st strand cDNA Synthesis Kit (Takara) and iTaq Universal SYBR Green Supermix (Bio-Rad). Real-time primers targeting NOX1, NOX2, NOX3, NOX4, NOX5, DUOX1, DUOX2, p22phox, p47phox, p67phox and actin were synthesized from Integrated DNA Technologies (IDT). The Sequence of primers are listed in Key Resources Table.