Validation of Cyclin D1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Cyclin D1 antibody (R020266, 1:2,000) and analyzed using BD flow cytometer.

Immunocytochemical staining of HeLa cells using Cyclin D1 antibody (R020266, 1:1,000), Top panel: wild-type (WT); Bottom panal: Cyclin D1 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Cyclin D1 was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm. Permeabilization: Triton.

Flow cytometric analysis of Cyclin D1 expression in C2C12 cells using Cyclin D1 antibody (R020266, 1:2,000). Green, isotype control; red, Cyclin D1.

Immunocytochemical staining of C2C12 cells with Cyclin D1 antibody (R020266, 1:1,000). Nuclei were stained blue with DAPI; Cyclin D1 was stained magenta with Alexa Fluor® 647. Images were taken using Leica stellaris 5. Protein abundance based on laser Intensity and smart gain: High. Scale bar: 20 μm.

Western blotting analysis using Cyclin D1 antibody (R020266). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Cyclin D1 antibody (R020266, 1:10,000) and HRP-conjugated goat anti rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using Cyclin D1 antibody (R020266). Cyclin D1 expression in wild type (WT) and Cyclin D1 shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with Cyclin D1 antibody (R020266, 1:10,000) and HRP-conjugated goat anti rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.