Flow cytometric analysis of LAT expression in HT-1080 cells using LAT antibody (R020807, 1:2,000). Green, isotype control; red, LAT.

Immunocytochemical staining of HT-1080 cells with LAT antibody(R020807, 1:1,000) . Nuclei were stained blue with DAPI; LAT 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 LAT antibody (R020807). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with LAT antibody (R020807, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit. LAT, linker for activation of T cells.

Western blotting analysis using LAT antibody (R020807). LAT expression in wild type (WT) and LAT shRNA knockdown (KD) HT-1080 cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with LAT antibody (R020807, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Validation of LAT knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with LAT antibody (R020807, 1:2,000) and analyzed using BD flow cytometer.