Fig. 1: Ribosome profiling identifies a microprotein encoded by a uORF in the 5′UTR of Lin28b. | Nature Communications

Fig. 1: Ribosome profiling identifies a microprotein encoded by a uORF in the 5′UTR of Lin28b.

From: Microprotein PLUM encoded by Lin28b uORF is a cytoplasmic determinant of pluripotency and embryonic development

Fig. 1: Ribosome profiling identifies a microprotein encoded by a uORF in the 5′UTR of Lin28b.

a Schematic representation of the Ribo-seq procedure in ESCs, EpiSCs and MEFs. b Ribosome profiling reveals widespread translation of non-canonical ORFs in ESCs, EpiSCs and MEFs. c Start codon usage in the identified ORFs. d Scatter plot showing the uORFs with significantly changed translation efficiency in EpiSCs compared with ESCs and MEF. e RPF (Ribosome protection fragment) densities in the 5′UTR and mORF of Lin28b in ESCs, EpiSCs and MEFs. f Amino acid sequence of pluripotency-associated Lin28b uORF-encoded microprotein (PLUM) from different species. g Left: Schematic representation of the expression constructs. Right: immunoblot analysis of translation of uORF in the 5′UTR of Lin28b in HEK293 cells. h Immunoblot analysis of endogenous PLUM expression in ESCs, EpiSCs, MEFs and PLUM overexpressing control (OE PLUM) samples. i Representative unique peptide spectrum and sequence of PLUM identified by LC-MS. j Immunofluorescence of PLUM-Flag in EpiSCs. Scale bar, 5 µm. k Immunoblot analysis of endogenous PLUM in nuclear and cytoplasmic fractions of EpiSCs. The experiments in g, h, j, k were repeated independently three times with similar results. Representative data are shown.

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