Figure 6

The XMRV pol sequence contributes a modest extent to steady-state level and nucleocytoplasmic export of gag mRNA. HEK293T cells were transfected with the indicated XMRV constructs. 48 hrs later, RNA from total (T) and cytoplasmic (C) fractions was collected. (A) XMRV mRNA was detected by northern blot with a radiolabeled gag mRNA probe. (B) The total cellular and cytoplasmic fractions used in northern blots were subjected to western blot with anti-HSP90 and anti-Histone H3 antibodies to monitor potential contamination of cytoplasmic preparations with nuclear contents. (C) Phosphorimager quantification of XMRV mRNA. Quantification was accomplished by detecting the bands and subtracting the background for each lane. This experiment - from transfection to northern blot quantitation - was repeated on three occasions with comparable results (see text). A representative experiment is shown.