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Free U4 accumulated in Δ25 and Δ30,20 cells. Total RNA was isolated from wild-type, Δ25, or Δ30,20 cells grown at 25°C or at 25°C, then shifted to 37°C for 2 h. (A) RNA was fractionated on a 5% nondenaturing polyacrylamide gel for Northern blotting probed sequentially with U4 and U6. (B) RNA was fractionated on a 5% polyacrylamide/8 M urea gel for Northern blotting and probed with U1, U2, U4, U5, and U6.
