Western blot analysis: Cell lysates of mouse brain and 293T (50 ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human DACT3 (1:250). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.

Paraffin embedded sections of human gastric cancer tissue were incubated with anti-human DACT3 (1:50) for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB)

Immunohistochemistry: Paraffin embedded sections of human gastric cancer tissue were incubated with anti - human DACT3 ( 1 : 50 ) for 2 hours at room temperature. Antigen retrieval was performed in 0.1M sodium citrate buffer and detected using Diaminobenzidine (DAB).