Fig. 6.

The expression of WEE1 was regulated by LINC00470 via sponging miR-580-3p in glioma cells. Note: The competitive binding sites of LINC00470 and WEE1 with miR-580-3p (a). After U251 and SWO-38 cells were transfected with pcDNA3.1-LINC00470 or sh-LINC00470 or their negative controls, the expression of miR-580-3p was examined by qRT-PCR (b). The expression of WEE1 was evaluated by qRT-PCR (c) and Western blotting (d), * P < 0.05, ** P < 0.01, compared to pcDNA3.1 group, # P < 0.05, ## P < 0.01, compared to sh-NC group. After U251 and SWO-38 cells were transfected with miR-580-3p mimic or miR-580-3p inhibitor or their negative controls, qRT-PCR evaluated the expression of miR-580-3p (e), LINC00470 (f) and WEE1 (g). Western blotting detected the expression of WEE1 (h), * P < 0.05, ** P < 0.01, *** P < 0.001, compared to mimic NC group, # P < 0.05, ## P < 0.01, compared to inhibitor NC group. Dual luciferase reporter assays were performed to assess the luciferase activity of U251 and SWO-38 cells (i–j), * P < 0.05, ** P < 0.01, compared to LINC00470-mut or WEE1-mut group. The binding of miR-580-3p with LINC00470 and WEE1 was detected by RIP (k), *** P < 0.001, compared to IgG group. NC, negative control; RIP, RNA immunoprecipitation