Non-transfected (–) and transfected (+) 293T whole cell extracts (30 μg) were separated by 12% SDS-PAGE, and the membrane was blotted with Nkx2.2 antibody (GTX133220) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.