Figure 2.
Nogo-(B) suppresses virus-induced interferon response. (A) Induction of IFN-β response of non-transformed and Ras-transformed NIH-3T3 cells was compared upon reovirus infection. (B) The efficient knockdown of Nogo gene was confirmed by western blot analysis. eGFP shRNA was used as a negative control. Relative protein levels (normalized to β-actin) were compared using ImageJ software (National Institutes of Health, Bethesda, Maryland, USA). (C and D) Effect of Nogo knockdown (KD) on reovirus-induced IFN response was analyzed in non-transformed (C) and Ras-transformed cells (D). Cells were infected with increasing reovirus doses (MOI of 200, 300, 400 based on their titers in L929 cells). (E) Non-transformed cells were transiently transfected 24 hr before viral infection with increasing amounts (0, 3, 6, 9 μg) of mouse Nogo-B expression plasmids. mRNAs were isolated at 24 hr post-infection and subjected to real-time q-PCR. Fold induction of IFN-β or ISG56 was normalized to GAPDH. eGFP shRNA was used as a negative control. Data are presented as mean values with error bars showing the standard deviations.