The Anti-interferon Activity of Conserved Viral dUTPase ORF54 is Essential for an Effective MHV-68 Infection
Figure 5
An ORF54-null virus infection promotes higher induction of interferon stimulated genes.
Bone marrow derived macrophages from wild type mice were infected at an MOI of 2. Equal infection was ensured by RT-PCR quantifying input viral genome copies from total infected cellular DNA 1 hour after virus was introduced to the culture. A) 24 hours post infection cells were harvested for immunoblot against murine IFIT2. The blot was stripped to demonstrate equal expression β-actin as a control. UI = uninfected; 54S = 54Stop; DM = 54DM. B) 24 hours post infection, RNA was harvested and reverse transcribed to cDNA. Transcripts for murine MX1, IFIT1, and IFIT3 were measured by real-time PCR. The fold induction was calculated by first normalizing each C(t) for the transcript to the actin C(t), and then comparing to uninfected samples.