Validation of ATG7 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) 293T cells were stained with ATG7 antibody (R021841, 1:2,000) and analyzed using BD flow cytometer.

Flow cytometric analysis of ATG7 expression in HT-1080 cells using ATG7 antibody (R021841, 1:2,000). Green, isotype control; red, ATG7.

Western blotting analysis using ATG7 antibody (R021841). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with ATG7 antibody (R021841, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.

Western blotting analysis using ATG7 antibody (R021841). ATG7 expression in wild type (WT) and ATG7 shRNA knockdown (KD) 293T cells with 20 μg of total cell lysates. β-Tubulin serves as a loading control. The blot was incubated with ATG7 antibody (R021841, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (1:20,000) respectively. Image was developed using ECL Substrate Kit.