Fig. 6

Effect of TNFα and IFNβ on the expression of rAd in APCs. Peritoneal macrophages were transfected with rAd-GFP in the presence of 0–30 ng/ml of TNF-α (c, e, f), 0–100 ng/ml IFN-β (a, e, f), or in the presence of combinations of 0–100 ng/ ml IFN-β and 10 ng/ml TNF-α (b), 0–30 ng/ml TNF-α and 10 ng/ml IFN-β (d). After 24 h, the efficacy of GFP protein synthesis (a-d), the level of transcription of the GFP (e) gene mRNA and the amount of viral DNA penetrated into the cells upon transfection (f) were analyzed. a-d GFP fluorescence in cells was detected by cytometry, mRNA (e) and DNA (f) from the lysates of DCs were extracted, RNA was subjected to a reverse transcription reaction; the obtained cDNA (e) and total cellular DNA (f) were used as a template for quantitative PCR with primers specific for GFP, GAPDH and β-actin genes. The expression values of GFP gene were normalized with the expression of β–actin or GAPDH genes. Values of GFP mRNA and DNA were normalized to the control values (No rAd). Shown are M ± SD, statistically significant (p < 0.05) differences are indicated by asterisks