Western blot analysis was performed on whole cell extracts (30 µg lysate) of MCF-7 (Lane1), A-431 (Lane 2), HT-29 (Lane 3), A549 (Lane 4), K-562 (lane 5), MDA-MB-231 (Lane 6), Colo 205 (Lane 7), HeLa (Lane 8), Caco2 (Lane 9) and Jurkat (Lane 10). The blots were probed with WNT4 Recombinant Rabbit Monoclonal Antibody (Product # 701857, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 40 kDa band corresponding to WNT4 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex®NuPAGE®4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).