Western blotting analysis using PTPN11 antibody (R020798). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with PTPN11 antibody (R020798, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit. PTPN11, protein tyrosine phosphatase non-receptor type 11.

Western blotting analysis using PTPN11 antibody (R020798). PTPN11 expression in wild type (WT) and PTPN11 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with PTPN11 antibody (R020798, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Flow cytometric analysis of PTPN11 expression in HepG2 cells using PTPN11 antibody (R020798, 1:2,000). Green, isotype control; red,PTPN11.

Validation of PTPN11 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with PTPN11 antibody (R020798, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HepG2 cells with PTPN11 protein antibody(R020798, 1:1,000). Nuclei were stained blue with DAPI; PTPN11 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: Medium. Scale bar, 20 μm.