Western blotting analysis using MAP2K3 antibody (R021751). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with MAP2K3 antibody (R021751, 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 MAP2K3 antibody (R021751). MAP2K3 expression in wild type (WT) and MAP2K3 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with MAP2K3 antibody (R021751, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Validation of MAP2K3 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with MAP2K3 antibody (R021751, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using MAP2K3 antibody (R021751, 1:1,000), Top panel: wild-type (WT); Bottom panal: MAP2K3 shRNA knockdown (KD). Nuclei were stained blue with DAPI; MAP2K3 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

Flow cytometric analysis of MAP2K3 expression in C2C12 cells using MAP2K3 antibody (R021751, 1:2,000). Green, isotype control; red, MAP2K3.

Immunocytochemical staining of C2C12 cells with MAP2K3 antibody (R021751, 1:1,000). Nuclei were stained blue with DAPI; MAP2K3 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.