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. 2015 May;21(5):1031–1041. doi: 10.1261/rna.047845.114

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© 2015 Rodgers et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 4. — SNAP-SiMPull analysis of U6 snRNP components. (A) Illustration of a SNAP-SiMPull experiment involving colocalization of DY649-labeled Prp24-SNAP_AP and DY549-labeled Lsm8-CLIP. (B) Representative microscopic fields of view from a U6 SNAP-SiMPull experiment. Lsm8-CLIP and Prp24-SNAP_AP were imaged simultaneously using a dual-view apparatus either in the presence or absence of ATP (left and center) or in the presence of ATP and after RNase H-directed cleavage of the U6 snRNA (right). Scale bar is 10 μm and all images are of identical magnification. (C) Quantification of colocalization of Prp24-SNAP_AP and Lsm8-CLIP from fields of view of identical dimensions. ATP causes a large increase in the extent of colocalization, while all colocalization is lost after cleavage of the U6 snRNA. The Random Control bar represents the probability of a Prp24-SNAP_AP spot of fluorescence colocalizing with a Lsm8-CLIP spot of fluorescence in the absence of genuine complexes being present. Error bars represent the standard error of the mean derived from three independent experiments.