Fig. 5

The microprotein TPM3P9 binds to RBM4 and suppresses TCF7L2 exon skipping. a Diagram showing the workflow of RNA pulldown combined with high-resolution mass spectrometry to identify proteins binding to TCF7L2 pre-mRNA. The diagram was edited using Adobe Illustrator. b Coomassie blue staining showing TCF7L2 pre-mRNA pulled down with RNA probes. The black arrows represent the lanes with significant differences. c Venn diagram showing a total of 65 proteins potentially interacting with TCF7L2 pre-mRNA. d Diagram showing the workflow of Co-IP combined with high-resolution mass spectrometry to identify TPM3P9-interacting proteins. The diagram was edited using Adobe Illustrator. e Silver staining showing proteins specifically bind to TPM3P9; specific bands are highlighted with red arrows. f Venn diagram showing 23 proteins potentially interacting with both TPM3P9 and the TCF7L2 pre-mRNA. g A PPI network of these proteins potentially interacting with both TPM3P9 and the TCF7L2 pre-mRNA was constructed using Cytoscape software. h The PPI network of the five interaction partners closely associated with TPM3P9 and the TCF7L2 pre-mRNA. i RNA pulldown with the TCF7L2 pre-mRNA probe was performed to validate the interaction between TCF7L2 pre-mRNA and RBM4. j Co-IP assays using an anti-Flag antibody were performed to detect the interaction between TPM3P9 and RBM4 in ACHN and 786-O cells. k Co-IP assays using an anti-GFP antibody were performed to detect the interaction between TPM3P9 and RBM4 in ACHN and 786-O cells expressing TPM3P9-Flag