Western blot was performed using Anti-SET Polyclonal Antibody (Product # PA5-78163) and a 38 kDa corresponding to SET was observed across cell lines tested. The expression of SET is low in MCF 10A when compared to MCF-7 and MDA-MB-231 as it is overexpressed in breast cancers. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), MCF-7 (Lane 2), MDA-MB-231 (Lane 3), PC-12 (Lane 4), NIH/3T3 (Lane 5) and MCF 10A (Lane 6) 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 (1:3000 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunofluorescent analysis of SET in HeLa cells. Samples were treated with 4% paraformaldehyde at RT for 15 min and incubated with SET polyclonal antibody (Product # PA5-78163) using a dilution of 1:500, followed by alpha tubulin (red) and Hoechst at a dilution of 1:1000.