Immunocytochemical staining of HepG2 cells with PDPK1 antibody (R020788, 1:1,000). Nuclei were stained blue with DAPI; PDPK1 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 PDPK1 antibody (R020788). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with PDPK1 antibody (R020788, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit. PDPK1, 3-phosphoinositide dependent protein kinase 1.

Western blotting analysis using PDPK1 antibody (R020788). PDPK1 expression in wild type (WT) and PDPK1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with PDPK1 antibody (R020788, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Validation of PDPK1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with PDPK1 antibody (R020788, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of PDPK1 expression in HepG2 cells using PDPK1 antibody (R020788, 1:2,000). Green, isotype control; red, PDPK1.