FIG. 3.

Reporter vRNAs are comparably and independently expressed in 293T cells. (A) RPA of reporter-specific negative-sense RNA (vRNA) in 293T cells transfected 48 h previously with 1 μg of each indicated reporter plasmid along with the 17-plasmid assembly system. (B) Reporter expression in 293T cells 48 h after transfection with 1 μg of each indicated reporter type either alone (alone) or in combination with an NA- or NP-based competitor plasmid encoding RFP (+ comp). Levels of vRNA were assayed by RPA with a probe that detects GFP and YFP but not RFP sequences and were quantified by phosphorimaging. Each of the five reporters was tested alone (in quadruplicate) and with each of the two competitors (in duplicate); data shown are mean ± standard error of the mean, pooled for homologous reporters. At right, titration of one representative cellular RNA sample to determine the sensitivity of the assay.