Western blot was performed using Anti-MYL9 Polyclonal Antibody (Product # PA5-17624) and a 20 kDa band corresponding to Myosin regulatory light polypeptide 9 was observed across cell lines and tissue extract tested and not in Daudi, Mouse Skeletal Muscle and Mouse Liver which is reported to be negative. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), Caco-2 (Lane 2), U-2 OS (Lane 3), Jurkat (Lane 4), MDA-MB-231 (Lane 5), C2C12 (Lane 6), Daudi (Lane 7) and tissue extracts Mouse Skeletal Muscle (Lane 8), Mouse Liver (Lane 9) and Mouse Urinary Bladder (Lane 10) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).