Skip to main content
. Author manuscript; available in PMC: 2020 Mar 4.
Published in final edited form as: Nature. 2019 Sep 4;573(7774):375–380. doi: 10.1038/s41586-019-1523-6

Fig. 1. In vitro assembled E complex is functional.

Fig. 1.

(a) The assembled E complex (with or without DNA oligo-directed RNase H treatment to cleave between the 5’ ss and BPS) is purified using the MS2 tag on pre-mRNA and its protein components shown. (b) Yeast splicing extract with or without U1 snRNA depletion is incubated with in vitro transcribed M3-Act1 or E complex assembled on M3-Act1 in the presence or absence of ATP or excess Act1-M3 (top gel). The splicing outcome is monitored using RT-PCR with primers located in the MS2 binding site region and exon 2 of M3-Act1. The middle and bottom gels demonstrate levels of U1 and U2 snRNA in each sample. Experiments in Fig. 1 were repeated two additional times with similar results. For all gel source data in this paper, see Supplementary Figure 1.