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. 2021 Jun 15;12:3646. doi: 10.1038/s41467-021-23934-y

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 7 — a TSSC4 downregulation does not change the U5 snRNP protein composition. Co-immunoprecipitation of U5-specific proteins with PRPF8-GFP was assayed after TSSC4 knockdown by western blotting. The positions of molecular weight markers are shown, the marker labeled by asterisk applies only to the GFP sample in the bottom combined GFP panel. b Sedimentation of U5-specific proteins is altered after TSSC4 knockdown and all three U5 markers migrate in lighter fractions than in control extracts. The positions of molecular weight markers are shown. a, b The experiments were performed three times with similar results. Uncropped and unprocessed scans are provided in the Source Data file. c Splicing efficiency is not globally reduced after TSSC4 downregulation. Total RNA was isolated, reverse transcribed, and analyzed by qPCR. The knockdown of PRPF8 served as a positive control. Mean and SEM from at least three biologically independent experiments are shown (in each experiment reverse transcription was performed in two technical replicates and the average value used for calculations). Statistical significance was determined by the paired two-tailed T test, P values are shown. Values for graphs are provided in the Source Data file. See also Supplementary Fig. 4 for knockdown efficiency.