Overexpression of CXCL12 rescues the EMT and migration abilities that were suppressed by GSK-J4. U-251MG and U-87MG ATCC cells were treated with GSK-J4 alone or GSK-J4 following the ectopic expression of CXCL12. (A) mRNA expression levels of CXCL12 and EMT-associated genes (E-cadherin, N-cadherin and fibronectin) in U-251MG (upper panel) and U-87MG ATCC (lower panel) cells. (B) Protein expression levels of JMJD3, E-cadherin, N-cadherin, CXCL12, CXCR4 and H3K27me3 were determined by western blotting in U-251MG and U-87MG ATCC cells treated with GSK-J4 or GSK-J4 and CXCL12 overexpression. (C) Wound healing assay to evaluate the migration ability of U-251MG and U-87MG ATCC cells treated with GSK-J4 or GSK-J4 and CXCL12 overexpression. Magnification, ×10. (D) Statistical results for the interval distance of the wound healing assay. (E) Representative DAPI staining images and (F) statistical results from the Transwell assay of the vector control, GSK-J4 treated and GSK-J4 plus CXCL12 overexpressing U-251MG and U-87MG ATCC cell groups. Scale bar, 200 µm. *P<0.05, **P<0.01. ATCC, American Type Culture Collection; CXCL12, C-X-C motif chemokine ligand 12; EMT, epithelial-mesenchymal transition; H3K27me3, histone H3 lysine 27 trimethyl.