Western blot was performed using Anti-PTPN2 Polyclonal Antibody (Product # PA5-80959) and 48 kDa, 45 kDa, and 40kDa bands corresponding to PTPN2 (Tyrosine-protein phosphatase non-receptor type 2) was observed across cell lines tested along with uncharacterized bands at ~80 kDa and 110 kDa. Whole cell extracts (30 µg lysate) of K-562 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), Caco-2 (Lane 4), MCF7 (Lane 5) and BJ (Lane 6) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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, 1:10000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunofluorescence analysis of Tyrosine-protein phosphatase non-receptor type 2 was performed using 70% confluent log phase K-562 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with PTPN2 Polyclonal Antibody (Product # PA5-80959) at 1:100 in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2500 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at __21 magnification.