Wild-type (WT) and ZAK knockout (KO) HeLa cell extracts (30 μg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with ZAK antibody [N3C2], Internal (GTX103317) 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.