Fig. 5: NUCKS1 promotes osteosarcoma tumorigenesis and metastasis via elevating ASNS expression.

A ASNS was knocked down in 143B and MNNG/HOS cells. The expression levels of ASNS were detected by western blot. B, C 143B and MNNG/HOS cells with or without ASNS depletion were tested for cell growth in the colony formation assay. After 1 week, viable colonies were counted and are shown (B). Data are depicted as bar graphs (C). D, E The migration of the indicated cells was detected by Transwell assays. Representative images of crystal violet-stained culture plates are shown (D). Data are depicted as bar graphs (E). F ASNS was knocked down in 143B and MNNG/HOS cells with or without NUCKS1 overexpression. The expression levels of ASNS and NUCKS1 were detected by western blot. G, H Cell growth were assessed by the colony formation assay. I, J Cell migration was assessed by Transwell assay. K–M The indicated 143B cells (10⁶ cells per mouse) were injected subcutaneously into nude mice (n = 4). Representative images of xenograft tumors (K). The volume (L) and weight (M) of the tumors were calculated and analyzed. N–O The indicated143B cells (10⁶ cells per mouse) were injected intravenously into nude mice (n = 5 per group). HE staining are displayed (N). Each group of metastatic nodules was assessed (O). Data in C, E, H, J, L, M, and O were analyzed by Student’s t test, *p < 0.05, **p < 0.01, ***p < 0.001.