Figure 6.

PLRG1 expressed in E.coli reduces the effect of CDC5L peptides in splicing inhibition. Lane 1, input pre-mRNA; lane 2 (CTRL1), a splicing reaction without peptides. The lane marked CTRL2 represents a splicing reaction containing the CDC5L peptide CD12-3. The splicing reaction in lane 4 contained CD12-3 peptide preincubated with GST (8 µg) for 10 min before addition to the reaction, whereas the samples in lanes 5 and 6 are duplicate splicing reactions containing CD12-3 peptide preincubated with an equivalent amount of GST–PLRG1 (i.e. for ∼10 min at room temperature prior to addition to the reaction). The contents of the splicing reactions in lanes 7 and 8 (duplicates) are similar to those of lanes 5 and 6 except that GST–PLRG1 was added after the splicing reactions had been incubated for 50 min at 30°C. The contents of the splicing reactions in lanes 9 and 10 (duplicates) are similar to that of lane 4 except that GST was added to the reactions after the splicing reaction had been incubated for 50 min at 30°C. The symbols on the right of the panel represent the input RNA, splicing intermediates and products. Other bands in the figure have been caused by partial degradation of the RNA in the reactions.