Non-transfected (–) and transfected (+) unboiled 293T whole cell extracts (30 μg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with Klotho antibody [HL3706] (GTX641878) diluted at 1:5000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.

Mouse tissue extracts (50 μg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with Klotho antibody [HL3706] (GTX641878) diluted at 1:1000. The HRP-conjugated anti-rabbit IgG antibody (GTX213110-01) was used to detect the primary antibody.