To further confirm the role of NUCB2 in breast cancer, we treated the breast cancer cells with exogenous Nesfatin-1 protein at doses of 400 pg/mL or 1000 pg/mL, which was shown to be less toxic (Additional file 1: Fig. S3A), and conducted migration and invasion assays on stable clones with NUCB2 knockdown. The gene discussed is NUCB2; the disease is breast cancer.