Validation of Nemo like kinase knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HT-1080 cells were stained with Nemo like kinase antibody (R021058, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of Nemo like kinase expression in HeLa cells using Nemo like kinase antibody (R021058, 1:2,000). Green, isotype control; red, Nemo like kinase.

Immunocytochemical staining of HeLa cells with Nemo like kinase antibody (R021058, 1:1,000). Nuclei were stained blue with DAPI; Nemo like kinase 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.

Western blotting analysis using Nemo like kinase antibody (R021058). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Nemo like kinase antibody (R021058, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using Nemo like kinase antibody (R021058). Nemo like kinase expression in wild type (WT) and Nemo like kinase shRNA knockdown (KD) HT-1080 cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with Nemo like kinase antibody (R021058, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.