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. 2019 Dec 12;40(3):697–713. doi: 10.1161/ATVBAHA.119.313430

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© 2019 The Authors.

Arteriosclerosis, Thrombosis, and Vascular Biology is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited.

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Figure 4. — PELATON (plaque enriched lncRNA in atherosclerotic and inflammatory bowel macrophage regulation) does not encode a micropeptide. A, PELATON predicted open reading frame (ORF; blue) appended with a hemagglutinin (HA) tag (yellow) was inserted into pcDNA 3.1(+) plasmid for in vitro translation assay. LINC00948 was used as a positive control, known to encode the micropeptide MLN (myoregulin). RNA expression using customized primers spanning ORF/HA boundary confirming transcription into RNA from plasmids by (B) quantitative real-time polymerase chain reaction (PCR) and (C) agarose gel electrophoresis of PCR products. D, Western blot using HA-antibody to detect tagged peptide produced by in vitro translation. E, Prediction scores estimating probability of protein-coding potential. M blank+L blank denotes quantitative real-time PCR negative controls for MLN-HA+PELATON ORF-HA, respectively. CPAT indicates coding potential assessment tool; CPC2, coding potential calculator version 2; CSF, codon substituion frequencies; and PRIDE, proteomics identification database.