Figure 2

Effects of Tap inhibitors on MLV RNA levels. Transfections were performed with pMov9.1Δenv, a Mo-MLV molecular clone deleted of the HpaI segment (1.38Kb) in the env gene in order to prevent re-infection events and the inhibition conditions were as in Figure 1. At 12 h (A), 24 h (B) and 48 h (C), RNAs were extracted, DNAse treated, and analyzed by RT-qPCR with an MLV standard curve as described in [15]. Primers used are indicated in Figure 1A and their sequences are given in Additional file 1 Table S1. The RT reaction was performed using oligodT or specific antisense primer and followed by different specific qPCR amplifications. Controls were systematically performed to check for DNA contamination by running an RT reaction without enzyme. Background RNA copies measured in untransfected cells were removed from values and then normalized to the GAPDH mRNA control monitored with specific probes as in [16]. Real time PCR was performed with the FastStart SYBRGreen (Roche) on a RotorGene (Corbett Research). Ribosomal RNAs were loaded on agarose gel and detected with SybrGreen staining as controls (D).