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. 2016 Sep 20;6:33605. doi: 10.1038/srep33605

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(a) Schematic overview of the FLAG-tagged PPP1R12A construct. Black vertical lines denote the ends of the overlapping region with R12A-AS1. The full overlapping region is highlighted with the red line (b) Western blot analysis of the FLAG-tagged PPP1R12A in HEK293T cells, transfected with the R12A-AS1 and pPPP1R12A-F expression constructs, as indicated. (c,f) qRT-PCR analysis of PPP1R12A and R12A-AS1 RNAs, extracted from HEK293T cells, in panel (b,e). (d) Schematic overview of synthetic miniSINEUPs, containing a FRAM or an inverted SINEB2 element, respectively. The S-region represents the spacer sequence. The length of each sub-element is indicated below (nt). (e) Western blot analysis of GFP in HEK293T cells after transfection with the two miniSINEUPs, shown in panel (d). Lower panel shows mean fold change ± s.d. (f) RT-qPCR analysis of RNA levels. *(n = 3, mean + S.D., p < 0.05, One-Sample t-Test, vs empty vector).