Enhanced expression of reporter RNA by REV-A RU5 is not due to altered cytoplasmic accumulation
Taken from "RNA helicase A interacts with divergent lymphotropic retroviruses and promotes translation of human T-cell leukemia virus type 1"
Nucleic Acids Research 2007;35(8):2629-2642.
Published online 10 Apr 2007
© 2007 The Author(s)() Nucleoplasm and cytoplasm were effectively isolated for analysis of mRNA cytoplasmic accumulation. Here, 293 cells were transiently transfected with indicated REV-A or SNV PCE reporter plasmids for 48 h. Nucleoplasm and cytoplasm were isolated and aliquots were subjected to immunoblot with antiserum to nuclear protein histone H1 and cytoplasmic protein β-tubulin, and β-actin loading control. () Summary table of results of Gag ELISA and RNA quantification. 293 cells transfected with indicated reporter plasmids were seperated into nuculear and cytoplasmic fractions. p(A) + RNA was isolated from each fractions, subjected to reverse transcption and real-time PCR with gag and β-actin primers. Values represent gag copy number per nanogram standarized to β-actin copy number. Ratio of Gag protein production determined by ELISA relative to cytoplasmic gag RNA level.