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. 2004 Jun 17;23(13):2632–2640. doi: 10.1038/sj.emboj.7600270

Figure 4.

Figure 4

The 4 × CTE RNA export element does not enhance budding in murine cells by increasing intracellular Gag concentration or by altering splicing. (A) The induction of Gag budding from 3T3 cells transfected with GPV-4 × CTE (lanes 1–3) versus GPV-RRE+pcRev (lane 4) is not due to variations in intracellular Gag concentration. Cells were transfected either with a titration of GPV-4 × CTE (100%=2 μg pGPV-4 × CTE+1 μg pcDNA3.1, total DNA levels were maintained by the addition of pcDNA3.1) or 2 μg of GPV-RRE+1 μg pcRev, and the Gag proteins were analyzed as in Figure 1A. (B) The RRE and 4 × CTE RNA export elements do not lead to differences in splicing. Cytoplasmic RNA was isolated from 3T3 cells transfected with GPV-RRE or GPV-4 × CTE and analyzed by Northern blot with a probe specific for the gag gene. The difference in transcript sizes is due to the 441 bp difference in the length of the RRE compared to the 4 × CTE.