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. 2018 Aug 27;8:12901. doi: 10.1038/s41598-018-31078-1

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© The Author(s) 2018

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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The SRSF3 protein and the NEXT complex associated with the RNA exosome are functionally connected to degrade intronless mRNAs. (a) Western blot analysis showing protein depletion upon the indicated siRNA treatments. Anti-Tubulin antibody was used as a loading control. * Indicates non-specific bands. (b) mRNA stabilization by exosome depletion: RT-qPCR analysis of nuclear RNAs from HeLa cells transfected with the indicated siRNAs together with the Renilla luciferase and BDLF1 intronless constructs. proEXT1 PROMPT served as a positive control. Data are displayed as mean values normalized to the control siRNA and GAPDH mRNA as an internal control. Error bars represent the s.d. from three independent experiments (n = 3). (c) Western blotting analysis showing the overexpression of the Flag tagged-hMTR4 protein and depletion of the SRSF3 protein after siRNA treatment. Anti-Tubulin antibody was used as a loading control. (d) RT-qPCR analysis of nuclear RNAs from HeLa cells transfected with the indicated siRNAs together with the Renilla luciferase and BDLF1 intronless constructs. Data are displayed as mean values normalized to the control siRNA and GAPDH mRNA as an internal control. Error bars represent the s.d. from three independent experiments (n = 3). (e) Western blotting analysis showing expression of the NEXT components RBM7, ZCCHC8 and hMTR4 in HeLa cells transfected with either a control siRNA (lane 1), or a siRNA specific for SRSF1 (lane 2), SRSF3 (lane 3) or SRSF7 (lane 4). Anti-Tubulin antibody was used as a loading control. *Indicates non-specific bands.