MCMV replication is inhibited by IE-3 specific shRNA. a) Semi-quantitative RT-PCR (30 cycles) of total RNA from MCMV infected M2-10B4 cells at either 3 or 6 hours p.i. Cells were transfected with shRNA-1 or a control shRNA 24 hours prior to MCMV infection. b) Western blot of proteins expressed in cells infected with MCMV and treated with either shRNA-1 or control shRNA. c) Viral growth curve showing the effect of shRNA-1 (dotted line), or control shRNA (solid line) on MCMV growth in M2-10B4 cells infected with MCMV at day 0 at an m.o.i. of 2. Results are typical of three such experiments performed. ** Significantly different from control p < 0.01. (d). Northern blot of total RNA extracted from MCMV infected M2-10B4 cells at various times p.i. following treatment with either shRNA-2 or control shRNA.