To gain further insights into the functions of LRP6 ubiquitin modifications induced by Itch, we used a doxycycline inducible HA-Ubiquitin expressing human osteosarcoma cell line, U2-OS, in which the expression of endogenous ubiquitin was replaced with HA tagged wild type ubiquitin [35]. Here, LRP6 is linked to osteosarcoma.