Western blotting analysis using anti-Ythdf1 antibody (Cat#65668). Total cell lysates (30 μg) from various cell lines were loaded and separated by SDS-PAGE. The blot was incubated with anti-Ythdf1 antibody (Cat#65668, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Western blotting analysis using anti-ythdf1 antibody (Cat#65668). YTHDF1 expression in wild-type (WT) and YTHDF1 shRNA knockdown (KD) HeLa cells with 20 μg of total cell lysates. Hsp90 α serves as a loading control. The blot was incubated with anti-ythdf1 antibody (Cat#65668, 1:5,000) and HRP-conjugated goat anti-rabbit secondary antibody (Cat#201, 1:20,000) respectively. Image was developed using NaQ™ ECL Substrate Kit (Cat#716).
Validation of Ythdf1 knockdown using flow cytometry. Wild-type(WT, Blue) and knockdown(KD, Green) HeLa cells were stained with anti-Ythdf1 antibody (Cat#65668, 1:2,000) and analyzed using BD flow cytometer.
Immunocytochemical staining of HeLa cells using anti-Ythdf1 antibody (Cat#65668, 1:1,000), Top panel: wild-type (WT); Bottom panal: Ythdf1 shRNA knockdown (KD). Nuclei were stained blue with DAPI; Acyl-CoA dehydrogenase short chain was stained magenta with Alexa Fluor® 647. Scale bar, 20 μm.
