Western blotting analysis using optineurin antibody (R020378). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with optineurin antibody (R020378, 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 optineurin antibody (R020378). Optineurin expression in wild-type (WT) and optineurin (OPTN) shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with optineurin antibody (R020378, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Validation of Optineurin knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Optineurin antibody (R020378, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using Optineurin antibody (R020378, 1:1,000), Top panel: wild-type (WT); Bottom panal: Optineurin 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.