Eif3h

Co-immunoprecipitation (co-IP) and western blot were performed as previously^{1 (link)}. Briefly, FLAG-METTL3 expressing HeLa or H1299 cells was harvested and lysed using NET-2 buffer [50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1 mM phenylmethanesulfonylfluoride, 2 mM benzamidine, 1% NP-40] then the supernatant was subjected to IP using Anti-FLAG M2 Affinity Gel (Sigma-Aldrich). Where indicated, the affinity elute was subjected to SDS-PAGE followed by either Colloidal Coomassie blue staining or Western blotting. Bands were excised and subjected to Mass spectrometric sequencing as previously^{1 (link)}. The following antibodies were used for Western blotting: METTL3 (Proteintech, 15073-1-AP; Abcam, ab195352), β-actin (Abcam, ab8227), eIF3h (Abcam, ab60942) CBP80 (Gift from Dr. Yoon Ki Kim, Korea University), CTIF (Gift from Dr. Yoon Ki Kim, Korea University), eIF4E (Cell Signaling Technology, #2067), eIF3b (Santa Cruz Biotechnology,sc-16377), eIF4GI (Cell Signaling Technology, #2498), FLAG (Sigma, A8592), BRD4 (Abcam, ab128874), CD9 (Cell Signaling Technology, #13174), MGMT (Cell Signaling Technology, #2739), TIMP1 (Cell Signaling Technology, #8946) and FTO (Phosphosolution, 597-FTO).