Vasculogenic mimicry (VM) formation assay.

U251 cells (I) or C6 cells (III) transfected with negative control microRNA (NC-DP) were seeded onto 6-well plates. ECM Matrigel was dropped onto 18-mm glass coverlips in 6-well tissue culture plate and incubated at 37°C for 30 min and then seeded on the coated coverlips. The VM formation was assessed using an inverted microscope after growth for 24 h. II and IV represent the U251 and C6 network formation stained by PAS, respectively. U251 (V) and C6 (VI) cells were transfected with 26b-DP, the transfected cells were seeded on the coated coverlips after 24 h incubation. The VM formation was assessed using an inverted microscope after growth for another 24 h. The U251 and C6 cells tansfected with 26b-DP first, then EphA2 expression vector which not include the 3′UTR (pCMV6-AC-GFP-EphA2) was co-transfected after 24 h incubation, respectively. The VM formation in U251 (VI) and C6 (VIII) cells was assessed using an inverted microscope after growth for another 24 h.