Immunocytochemical staining of HAP1 cells using NELFB antibody (R021429, 1:1,000), Top panel: wild-type (WT); Bottom panal: NELFB shRNA knockdown (KD). Nuclei were stained blue with DAPI; Acyl-CoA dehydrogenase short chain was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.

Immunocytochemical staining of HepG2 cells with NELFB antibody (R021429, 1:1,000). Nuclei were stained blue with DAPI; NELFB 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 NELFB antibody (R021429). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with NELFB antibody (R021429, 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 NELFB antibody (R021429). NELFB expression in wild-type (WT) and NELFB shRNA knockdown (KD) HSHC cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with NELFB antibody (R021429, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Validation of NELFB knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HSHC cells were stained with NELFB antibody (R021429, 1:2,000) and analyzed using BD flow cytometer.