The ORF6 protein exerts a transcriptional effect on nsp8 expression. Vero E6 cells were co-transfected with 1 μg of pXJ40myc-nsp8 and different amount of pXJ3'-ORF6. For each sample, the total amount of DNA was normalized to 3 μg by addition of the empty vector, pXJ3'HA, when necessary. Total RNA was harvested 16 hours later, reverse transcribed and subjected to quantitative real-time Taqman PCR using primers and probes specific to nsp8 and cellular protein, GADPH. Relative quotients between the resulting Ct values for each sample were calculated using the ΔΔCt method. Similar results were obtained in three independent experiments, and a representative set of data is presented.