Western blot was performed using Anti-Apelin Receptor Polyclonal Antibody (Product # PA5-80343) and a 42 kDa band corresponding to Apelin receptor was observed across cell lines tested. Whole cell extracts (20 µg lysate) of Hep G2 (Lane 1), K-562 (Lane 2), PC-3 (Lane 3), Daudi (Lane 4) and Jurkat (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Jurkat expresses APLNR protein at lower levels as compared to Daudi. 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 (0.2 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).