Extended Data Fig. 3: METTL3 associates with translation initiation factors.
From: mRNA circularization by METTL3–eIF3h enhances translation and promotes oncogenesis
a, Deletion mutants of METTL3 were expressed in HeLa cell. The total-cell extracts (Input) and the cap-associated protein samples were analysed by western blotting using the indicated antibodies. b, Cap-association assay with METTL3 depletion. The total-cell extracts (Input) and the cap-bound protein samples were analysed by western blotting using the indicated antibodies. m7GpppG cap analogue was used for antagonizing cap-associating proteins binding to m7GTP-agarose. c, Same as b, except HeLa cells were transfected with CTIF, EIF3B or EIF4G1 siRNA. In a, b, blots are representative of two independently performed experiments with similar results. d–f, Mass spectrometry of Flag–METTL3 interacting proteins. d, Proteins that were co-immunopurified with Flag–METTL3 subjected to 4–12% Tris–glycine SDS–PAGE. Colloidal Coomassie blue staining was performed. e, Gene Ontology analysis of the identified proteins from mass spectrometry. One experiment was performed. Hypergeometric distribution (one-tail) with Bonferroni adjustment was used to determine enrichment statistical significance. f, Table showing the translation involving factors identified from mass spectrometry.
