Fig. 3

EWSR1 promotes the biogenesis of circNEIL3. A The binding sites of EWSR1 were predicted in the upstream and downstream region of NEIL3 pre-mRNA transcript using the starbase and circinteractome database. B Relative expression of EWSR1 in our local NBTs, LGG and GBM tissues detected by high-throughput sequencing. C CCK8 assays showing the proliferation ability of GBM cells transfected with sh-NC or sh-circNEIL3, n = 3. D Representative transwell migration and invasion assays showing the migration and invasion ability of GBM cells transfected with sh-NC or sh-circNEIL3. Quantification histogram represented relative cell numbers, n = 3, scale bar, 200um. E The correlation between circNEIL3 and EWSR1 expression in our local glioma high-throughput sequencing data (n = 39). F qRT-PCR assays showed the expression of circNEIL3 in GBM cells transfected with sh NC or sh EWSR1, n = 3. G Western blot assays showing the expression of EWSR1 under hypoxia treatment for 0, 6, 12, 24 h. H qRT-PCR assays showing the expression of circNEIL3 under hypoxia treatment for 0, 6, 12, 24 h, n = 3. I RIP and qRT-PCR assays were performed to verify the putative binding site of NEIL3 pre-mRNA with EWSR1 under normal and hypoxia environment, n = 3. (J) CCK8 assays, (K) Representative EDU assays showing the proliferation ability of GBM cells co-transfected with ov-NC, ov-EWSR1 and si-circNEIL3 as indicated. Quantification histogram represented relative cell numbers, n = 3, scale bar = 50um. L Representative transwell migration and invasion assays showing the migration and invasion ability of GBM cells co-transfected with ov-NC, ov-EWSR1 and si-circNEIL3 as indicated, Quantification histogram represented relative cell numbers, n = 3, scale bar, 200um. All data are presented as the means ± SD, and the ov-EWSR1 group is indicated as the control in (J-L), ns, P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001