Western blot analysis was performed on Whole cell extracts (30 µg lysate) of MCF-7 (Lane 1), MCF-7 treated with PMA (50 ng/mL for 48hrs) (Lane 2), T98G (Lane 3) and U-87 MG (Lane 4). The blots were probed with Anti-DUSP6 Rabbit Polyclonal Antibody (Product # 720405, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.2 µg/mL, 1:5000 dilution). A 42 kDa band corresponding to DUSP6 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0322BOX), 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).