Conditional overexpression of Dlx2 in neural crest cells

We obtained wnt1^cre mice from the Jackson laboratory. The Rosa26^{CAG-LSL-Dlx2−3xFlag} mouse was constructed by the Shanghai Model Organisms Center, Inc. (Shanghai, China). To generate wnt1^cre; Rosa26^Dlx2/- mice, which could specifically overexpress Dlx2 in neural crest cells, we mated wnt1^cre mice with Rosa26^{CAG-LSL-Dlx2−3xFlag} mice. Wildtype C57BL/6J mice were purchased from Shanghai Jihui Laboratory Animal Care Co. Ltd. (Shanghai, China). All mice were maintained under SPF conditions at the Animal Center of the Ninth People’s Hospital affiliated with Shanghai Jiao Tong University School of Medicine. The day of the appearance of a vaginal plug was defined as E0.5 in all timed pregnancies. Embryos at the E10.5 and E12.5 stages (12:00 h of the day when the vaginal plug was detected was counted as E0.5) and P0 pups were collected for subsequent experiments. All animal experiments were approved by the Animal Care and Usage Committee of the Ninth People’s Hospital affiliated to Shanghai Jiao Tong University School of medicine.