Western Blot was performed using Anti-TRIB3 Polyclonal Antibody (Product # PA5-15480) and a 40 kDa band corresponding to Tribbles homolog 3 was observed across cell lines and tissue tested. The protein expression was found to be upregulated in Hep G2 upon treatment with Tunicamycin and TGF beta 1 treatments. Nuclear enriched extracts (30 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with Tunicamycin (2 µg/mL for 12 hrs) (Lane 2), Hep G2 treated with TGF beta 1 (10 ng/mL for 12 hrs) (Lane 3), MDA-MB-231 (Lane 4), MCF7 (Lane 5), SW480 (Lane 6), SiHa (Lane 7), Mouse Liver (Lane 8) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) 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, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Additional uncharacterized band was also observed at ~45 kDa.