Fig. 3

MiR-5047 is a target of WAC-AS1. A. Prediction of target miRNAs potentially binding to WAC-AS1 was performed using ENCORI and miRDB. Expression of miRNAs was determined in control and WAC-AS1-transfected OS cells. B. Expression of miR-5047 was detected after transfecting WAC-AS1 or sh-WAC-AS1. C. Wild-type and mutant WAC-AS1 binding sites in miR-5047. Luciferase reporter assay detecting the binding of miR-5047 to WAC-AS1 in OS cells. D. RIP using anti-AGO2 antibody was performed to detect WAC-AS1 and miR-5047. E. Transwell assay performed to detect the migtaion of WAC-AS1- or miR-5047-transfected or control OS cells (scale bar, 50 μm). F. Western blotting to determine stemness-related proteins in control and indicated transfected cells. G. Changes in tumorsphere-forming ability of control and indicated transfected OS cells (scale bar, 50 μm). H. Percentage of CD133 + cells in WAC-AS1-overexpressing, SOX2-transfected or WAC-AS1-knockdown and control OS cells was determined by flow cytometry. I. Expression of miR-5047 was measured by qRT-PCR in OS tissues and normal tissues. J. Correlation between WAC-AS1 and miR-5047 expression in OS. Error bars represent three independent experiments, *p < 0.05, **p < 0.01, *** p < 0.001