Western blot was performed using Anti-FTO Recombinant Rabbit Monoclonal Antibody (Product # 703602) and a 58 kDa band corresponding to FTO was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of SH-SY5Y (Lane 1), U-2 OS (Lane 2), A549 (Lane 3) and THP-1 (Lane 4) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036) at a 1:5000 dilution. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).