Western blot was performed using Anti-ATG7 Polyclonal Antibody (Product # PA5-17216) and a 70 kDa band corresponding to ATG7 was observed across cell lines tested and increased upon Chloroquine treatment. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), HeLa treated with Chloroquine (50nM for 15 hr) (Lane 2), HCT 116 (Lane 3) and HCT 116 treated with Chloroquine (50nM for 15 hr) (Lane 4) were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX). 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).