Wild-type (WT) and 4E-BP1 (phospho Ser65) knockout (KO) HeLa cell extracts (30 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with 4E-BP1 (phospho Ser65) antibody (GTX133184) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.

Untreated (–) and treated (+) 293T whole cell extracts (60 μg) were separated by 15% SDS-PAGE, and the membrane was blotted with 4E-BP1 (phospho Ser65) antibody (GTX133184) diluted at 1:500.