Western blot was performed using Anti-IL-6 Monoclonal Antibody (6708) (Product # MA5-23698) and a 24 kDa band corresponding to IL-6 was observed across all cell treatments tested. Whole cell extracts (50 µg lysate) of HUVEC (Lane 1), HUVEC treated with LPS and Brefeldin A (0.5 µg/mL of LPS for 24 h and 300 ng/mL of Brefeldin A for last 20 h of LPS stimulation) (Lane 2), HaCaT (Lane 3), HaCaT transfected with Poly(I:C) and treated with PTI (0.67 µg/mL and 1x respectively, for 4 h) (Lane 4), HaCaT (Lane 5) and HaCaT transfected with Poly(dA:dT) and treated with PTI (0.67 µg/mL and 1x respectively, for 12h and last 4 h of stimulation) (Lane 6) 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 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005). A 24 kDa band corresponding to IL-6 was observed to be upregulated upon cell treatment.