Fig. 3

FAM289 promotes proliferation and migration of GBM cells dependent on the interaction with Galectin-1 protein. (A) Exogenous FAM289 interacts with Galectin-1. HEK293T cells were transfected with peGFP-FAM289 and Flag-tagged Galectin-1 plasmids. Co-IP experiment was performed using anti-Flag antibody. (B) Endogenous FAM289 interacts with plasmid-produced Galetin-1. Co-IP experiment was performed using anti-FAM289 antibody in U251 and U87-MG cells. IgG was used as the negative control. The immunoprecipitates were analyzed via western blotting with anti-FAM289 and anti-Galectin-1 antibodies. (C-H) The effect of FAM289/Galectin-1 interaction on the tumor-promoting functions of FAM289.SiRNAs specifically targeting Galectin-1 were transfected into U251 con and U251-FAM289^OE cells to knockdown endogenous Galectin-1.These cells’ proliferation and migrating capacity were analyzed by xCELLigence RTCA (C), scratch assay (D) and transwell assay (E). Galectin-1vectors were transfected into U87-MG con and U87-MG-FAM289 KD cells, and these cells’ proliferation and migrating capacity were analyzed by xCELLigence RTCA (F), scratch assay (G) and transwell assay (H). Each tracing represents an average of three parallel assessments. Scale bars. *P < 0.05, **P < 0.01 vs controls