Extended Data Fig. 1: MARTFQ screening reveals that NADP directly binds FTO.
From: NADP modulates RNA m6A methylation and adipogenesis via enhancing FTO activity
a, Schematic showing MARTFQ assay. b, 3D structure of FTO protein containing tryptophan residues. The data for the FTO 3D structure were derived from NCBI accession code: 3LFM (Protein Data Bank). Red dots represent tryptophan residues in FTO protein. c, Coomassie staining of in vitro purified FTO protein. d, 3D structure of FTO. The data of FTO 3D structure were derived from accession code: 3LFM (Protein Data Bank). N terminal: blue; C terminal: red; 2KG: green. e, Coomassie staining of in vitro purified 6XHis-FTO-NT and 6XHis-FTO-CT proteins. f, Quenching of the fluorescence of FTO-NT (left) and FTO-CT (right) proteins by NADPH at different molar ratios. g, The modified Stern–Volmer curves for estimating the K values of FTO-NT (18.5 x 103 M-1) and FTO-CT (30.2 x 103 M-1). h, Immunoblotting analyses of the cell lysates from HEK293 cells transfected with pCDNA3 Flag-FTO (WT), pCI-Neo Flag-FTO-NT or pCI-Neo Flag-FTO-CT plasmid. i,j, DARTS (i) and CETSA (j) analyses of the abilities of NADPH to bind FTO-NT and FTO-CT in HEK293 cells transfected with pCI-Neo Flag-FTO-NT or pCI-Neo Flag-FTO-CT plasmid. Immunoblotting was performed using anti-Flag antibody.
