Extended Data Fig. 3: METTL3 interacts with NUP93 at NPCs.
From: N6-adenosine methylation enhances nuclear mRNA export through METTL3 and NUP93
a,b, Reciprocal IP assays using anti-GFP (a) or anti-HA (b) antibodies in total lysates from HeLa cells exogenously expressing GFP-tagged NUP93 and HA-tagged METTL3. c, IP assays showing that the interaction of METTL3 and NUP93 is mediated by protein-protein contacts. Treatment with RNase A or DNase I during IP did not disrupt the interaction. d, In vitro GST pull-down assay demonstrating a direct interaction between METTL3 and NUP93. GST-NUP93 proteins (amino acids 197-552) were used. e, IP assays showing that the interaction of METTL3 and NUP93 occurs in multiple cell types. f, An in situ proximity ligation assay (PLA) showing interaction between METTL3 and NUP93 at the peripheral nuclear membranes in multiple cell types. g, An in situ PLA using transcription factors that translocate through nuclear pore complexes (MYC and ETS1), RNA polymerase II (RNAPII), or METTL14 as negative controls showed no detectable PLA signal with NUP93, supporting the specificity of the METTL3-NUP93 interaction. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Scale in (f, g) bar, 2 μm. All panels show representative images from three independent experiments. Source unprocessed gels and blots are provided in the Source Data.
