Western blot was performed using Anti-Myogenin Monoclonal Antibody (F5D), eBioscience™ (Product # 14-5643-82) and a 30 kDa band corresponding to MYOG was observed across cell lines except HEK-293 and NIH/3T3 which are reported to be negative and also increased upon 2% heat inactivated horse serum treatment. Modified whole cell extracts (1% SDS) (30 µg lysate) of C2C12 (Lane 1), C2C12 treated with 2% heat inactivated horse serum for 5days (Lane 2), RD (Lane 3), L6 (Lane 4), HEK-293 (Lane 5) and NIH/3T3 (Lane 6) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 (2.5 ug/ml) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).