Western Blot was performed using Anti-FTO Polyclonal Antibody (Product # PA5-89791) and a 58 and 58.2 kDa band corresponding to Alpha-ketoglutarate-dependent dioxygenase FTO was observed across cell lines and tissues tested . Nuclear enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), U-87 MG (Lane 2), Hep G2 (Lane 3), PC-3 (Lane 4), HeLa (Lane 5), MCF7 (Lane 6), Mouse Skeletal Muscle (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).Both isoforms around 58 k Da picked up signal as seen.