Validation of Tuberin knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with Tuberin antibody (R020248, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of Tuberin expression in HeLa cells using Tuberin antibody (R020248, 1:2,000). Green, isotype control; red, Tuberin.

Immunocytochemical staining of HeLa cells with Tuberin antibody (R020248, 1:1,000). Nuclei were stained blue with DAPI; Tuberin 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 Tuberin antibody (R020248). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with Tuberin antibody (R020248, 1:5000) and HRP-conjugated goat anti-rabbit secondary antibody (1:50,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using Tuberin antibody (R020248). Tuberin expression in wild type (WT) and Tuberin shRNA knockdown (KD) HeLa cells with 30 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with Tuberin antibody (R020248, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.