Mutations in RNAP II subunit RPB2 rescue exon inclusion. (A) Splicing of dyn2 U[−2]C. (B) Splicing of dyn2 A[2]G. All reactions were normalized to the E2+ signal before loading on the gel, so that levels of skipping can be evaluated by observing changes in the amount of the E2− signal. (C) Quantitation of dyn2 cDNA signals and calculation of alternative splicing ratios (±SEM; n = 3 for U[−2]C and n = 4 for A[2]G). Each dyn2 cDNA was measured and is expressed as the percentage of the total dyn2 cDNA signal in each lane. Measures of the statistical confidences of E2+, E2−, and E2−/E2+ values relative to the wild-type RPB2 strain were calculated. Student’s t test was applied to compare E2+, E2−, or E2−/E2+ quantities in mutant with the wild-type strain. P values <0.05 indicate that the differences between the compared values are unlikely to be due to sampling error.
