Certain mutations in the Jab1/MPN result in reduced levels of U4/U5/U6 tri-snRNP. Splicing extracts from mutant and wild-type strains were incubated at 25°C in the presence or the absence of 2 mM ATP. The reactions were resolved by native polyacrylamide gel electrophoresis, transferred to Hybond-N+ membranes, and sequentially probed for U6, U5, and U4 snRNAs. The position of U4/U5/U6 tri-snRNP, U4/U6 di-snRNP, and the free snRNP complexes are indicated to the left. The rightmost lane in all three panels was loaded on the left side of the gel and then repositioned using Adobe Photoshop. All digital rendering was done to the entire file prior to cutting and pasting that lane.
