Western blot was performed using Anti-Complement C3b Monoclonal Antibody (013III-1.16) (Product # MA1-82814) and a 187 kDa band corresponding to Complement C3 was observed in HepG2 in comparison to HEK-293 which is reported to be negative. A band around 120 kDa was also observed in Hep G2 treated with LPS and PTI which can be the processed form of complement C3. Whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with LPS (10 ng/mL for 18 hr) followed by protein transport inhibitor (1X for 4 hr) (Lane 2) and HEK-293 (Lane 3) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).