Western blot was performed using Anti-IL-6 Polyclonal Antibody (Product # PA1-26811) and a 24 kDa band corresponding to IL-6 was observed across to be upregulated upon treatment of HUVEC cells with LPS and Brefeldin A. Whole cell extracts (50 µg lysate) of HUVEC (Lane 1) and HUVEC treated with LPS and Brefeldin A (0.5 µg/mL LPS for 24 h and 300 ng/mL Brefeldin A for last 20 h of LPS stimulation) (Lane 2) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).Expression of IL-6 was observed to be to be upregulated upon treatment of HUVEC cells with LPS and Brefeldin A, HaCaT cells with Poly(I:C) and PTI, and Poly(dA:dT) and PTI.