Figure 3.

Inhibition of RNA export in oocytes. (A) A mixture of radioactively labeled m⁷G-U1ΔD, U6, and tRNA was injected into the nucleus of oocytes in the absence (−) or presence of unlabeled, uncapped competitor RNA (XAP1, V_TL). After 1 h oocytes were separated into nuclear (N) and cytoplasmic (C) fractions, and extracted RNA was analyzed on a denaturing acrylamide gel (the T7htRNA appears to be longer once transported to the cytoplasm; apparently nucleotides are added to the 3′ end before or during the translocation step). (B) As above. In addition, RNA was extracted from total oocytes immediately after injection (T₀) and after 1 h (T₆₀). (C) Labeled ApppG-capped Ad46 was preincubated in the presence (+) or absence (−) of Rev protein, mixed with labeled U6, and injected into the nucleus of oocytes either alone (no comp.) or together with unlabeled, uncapped competitor (XAP1, V_GG). Oocytes were separated after 45 min into nuclear (N) and cytoplasmic (C) fractions, and extracted RNA was analyzed.