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. Author manuscript; available in PMC: 2021 Dec 1.
Published in final edited form as: Nat Biotechnol. 2021 Jan 28;39(6):697–704. doi: 10.1038/s41587-020-00806-2

Figure 1: Identification of translated unannotated or unstudied open reading frames.

Figure 1:

a) A schematic overview of the research project. b) The experimental set-up for in vitro detection of protein translation by transfection of V5-tagged cDNAs into HEK293T cells followed by in-cell western blotting. c) In-cell western blot signal for each ORF. Values are the average of three replicates. d) Immunoblot correlates for three ORFs identified by in-cell western blotting, marked in panel c. Results were repeated in three independent experiments. e) An overview of biological support for translation of a subset of ORFs. f) Subgroup analyses of ORF biological features demonstrating fractions of ORFs supported by ectopic V5 translation assays, mass spectrometry or both. g) The fraction of ORFs supported by evidence of translation across major epochs in evolutionary time. Evidence of translation shown as the fraction of ORFs with V5 western blot signal, endogenous mass spectrometry peptides, and the summation of both.