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. Author manuscript; available in PMC: 2014 Feb 21.
Published in final edited form as: Mol Cell. 2011 Mar 4;41(5):579–588. doi: 10.1016/j.molcel.2011.02.012

Figure 2. U1 snRNP protects a much longer region of the pre-mRNA under PTB-dependent repression.

Figure 2

A) The extent of protection of the labeled c-src RNAs was determined using oligonucleotide mediated RNase H cleavage. Site specifically labeled wildtype RNAs were incubated in HeLa extract (lanes 1–8) and WERI extract (lanes 9–16). After incubation, reactions were treated with micrococcal nuclease (lanes 2–8 and 10–16). The protected fragments were extracted using PCA and ethanol precipitated. The protected RNAs were then subject to RNase H cleavage in presence of 15 nucleotide long oligonucleotides as indicated. After RNase H treatment, the RNAs were separated using urea-PAGE and visualized by phosphorimaging. Oligos with complimentarity offset from those shown here gave equivalent results in both HeLa and WERI extracts (see Figure S2). B) Sequence of the N1 exon containing pre-mRNA. Positions of DNA oligonucleotides used for RNase H cleavage are indicated above the sequence. The lines below the sequence indicate the boundaries of nuclease protection in HeLa and WERI extracts.