Fig. 4. The effect of repression of TbMP81 expression on in vitro pre-cleaved U insertion and deletion editing. Editing complexes were immunoprecipitated with MAb P1H3-D7 (specific for TbMP63) from total cell lysates that were grown in the absence of tet (no RNAi) or in the presence of tet (RNAi). (A) Pre-cleaved insertion editing by the immunoprecipitates. Products from PF immunoprecipitates with two added Us (+2U), edited by the insertion of two Us (+2), or the ligated input (0) RNAs are indicated in the top panel. (B) Pre-cleaved deletion editing by the immunoprecipitates. Products with 4 Us removed (–4U), edited by deletion of 4 Us (–4), and the ligated input (–0U) RNAs are indicated in the top panel. The percentage of +2 or –4 edited and +2U addition or –4 removal products as a function of total input RNA in PF after 4 days and in BF after 3 days, as well as the percentage inhibition upon RNAi induction (+tet), are indicated in the lower panels. Similar experiments were performed with BF immunoprecipitates. Pre-cleaved insertion and deletion editing experiments were repeated four and three times, using PF and BF immunoprecipitates, respectively.