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. 2018 Sep 26;46(22):12022–12039. doi: 10.1093/nar/gky862

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© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — RNA binding of RRM is needed for the enhancement of SLAP provided by CstF-77. (A) Schematic illustration of point mutants in sites I, II and III of the RRM of MCP-CstF-64. (B) SLAP of the wild- type and I, II and III mutants of the RRM of CstF-64 alone and co-expressed with CstF-77-Myc. Western blots were performed to show expression of MCP-CstF-64 constructs and CstF-77-Myc. β-tubulin was used as a loading control. (C) CLIP experiment verifying that site I, II and III MCP-CstF-64 mutants do not bind or bind minimally to RNA (bottom). Western blot of the immunoprecipitated samples is also shown (top). (D) Immunohistochemistry staining of site I, II and III MCP-CstF-64 mutants. Site I and II localize to the nucleus when expressed alone (green). Site III demonstrates a cytoplasmic localization when expressed alone (green). All MCP-CstF-64 mutants locate to the nucleus when co-expressed with CstF-77-Myc (red).