Fig. 5. (A) Commitment complex analysis of the cs mutant. Extracts from cells grown at permissive (30°C) or non-permissive (16°C) temperature were incubated with labeled pre-mRNA for 30 min at 30 or 16°C in the absence of ATP. Reactions containing extracts from the mutant strain (cs) or the wild-type control (WT) are indicated above each lane. A control reaction was performed with the wild-type extract and a labeled pre-mRNA lacking the branchpoint sequence (C) to obtain a marker for the migration of CC1. (B) (Pre)-spliceosome analysis of cs mutant. Extracts from cells grown at permissive (30°C) or non-permissive (16°C) temperature were incubated with labeled pre-mRNA for 30 min at 30, 16 or 14°C in the presence of ATP. Labeling of the gel is as in (A).