Western blot was performed using VASP Polyclonal Antibody (Product # PA5-80212) and a 50kDa band corresponding to VASP was observed across cell lines along with an uncharacterized band (*) at ~45 kDa. Whole cell extracts (30 µg lysate) of HEL 92.1.7 (Lane 1), A431 (Lane 2), HEK293 (Lane 3), HeLa (Lane 4) and SHSY5Y (Lane 5) were electrophoresed using NuPAGE® 4-12 % Bis-Tris 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 (1:1000 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).

Immunohistochemistry analysis of VASP on paraffin-embedded human mammary cancer tissue. Sample was incubated with VASP polyclonal antibody (Product# PA5-80212) with a dilution of 1 µg/mL, and developed by Streptavidin-Biotin-Complex (SABC) method.