HepG2 whole cell extract and conditioned medium (30 μg) were separated by 12% SDS-PAGE, and the membrane was blotted with Shh antibody (GTX134453) 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.