Validation of Bmi1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Bmi1 antibody (R020267, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of Bmi1 expression in HepG2 cells using Bmi1 antibody (R020267, 1:2,000). Green, isotype control; red, Bmi1.

Immunocytochemical staining of HeLa cells with Bmi1 antibody (R020267, 1:1,000). Nuclei were stained blue with DAPI; Bmi1 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 Bmi1 antibody (R020267). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Bmi1 antibody (R020267,1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using Bmi1 antibody (R020267). Bmi1 expression in wild type (WT) and Bmi1 shRNA knockdown (KD) 293T cells with 30 μg of total cell lysates. GAPDH serves as a loading control. The blot was incubated with Bmi1 antibody (R020267,1:5,000) and HRP-conjugated goat anti rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.