Western blot was performed using Anti-TNF alpha Polyclonal Antibody (Product # PA5-19810) and a 25kDa band corresponding to TNF-Alpha was detected in lipopolysaccharide treated macrophages (differentiated from both THP-1 and U-937 and RAW 264.7 macrophage). Whole cell extracts (30 µg lysate) of untreated macrophages differentiated from U-937 (Lane 1), treated with PTI (Lane 2), stimulated with LPS (Lane 3), stimulated with LPS and subsequently treated with PTI (Lane 4), untreated RAW 264.7 (Lane 5), stimulated with LPS (Lane 6), stimulated with LPS and subsequently treated with PTI (Lane 7) in Fig. a, and untreated, differentiated macrophage from THP-1 (Lane 1), differentiated macrophage treated with PTI (Lane 2), THP-1 macrophage stimulated with LPS (Lane 3), THP-1 macrophage stimulated with LPS and then treated with PTI (Lane 4) in Fig. b were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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 (0.9 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:8000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).