Western blot was performed using Phospho-STAT6 (Tyr641) Polyclonal Antibody (Product # PA5-17902) and a 95 kDa band corresponding to phospho-STAT6 was observed in ACHN cells on treatment with IL-4. Whole cell extracts (30 µg lysate) of ACHN (untreated) (Lane 1), ACHN treated with IL-4 (100 ng/mL for 15 minutes) (Lane 2), ACHN treated with IL-4 (100 ng/mL for 30 minutes) (Lane 3), LNCaP (untreated) (Lane 4), LNCaP treated with IL-4 (100 ng/mL for 30 minutes) (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX), 10 well. 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) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescentdetection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).