Fig. 5

MIR17HG negatively regulates the eexpression of miR-153(mir-377) and miR-153(mir-377) is involved in MIR17HG-induced permeability regulation of glioma-conditioned normal BBB and tight junction-related proteins. (a) miR-153 (miR-377) expression levels in MIR17HG knockdown GECs on quantitative real-time PCR. Data represent the mean ± SD (n = 3, each). **P < 0.01 vs. shMIR17HG-NC groups. (b) Relative luciferase activity in HEK293T cells co-transfected with pmirGLO-MIR17HG-Wt (pmirGLO-MIR17HG-Mut) and agomir-153 (agomir-377). Data represent the mean ± SD (n = 3, each). **P < 0.01 vs. the MIR17HG-Wt + miR-153 (+) group, *P < 0.05 vs. the MIR17HG-Wt + miR-377 (+) group. (c) RNA immunoprecipitation assay performed with normal mouse IgG or anti-Ago2. Relative enrichment of MIR17HG and miR-153 (miR-377) was determined on quantitative real-time PCR. Data represent the mean ± SD (n = 3, each). **P < 0.01 vs. the IgG group, #P < 0.05 and △P < 0.05 vs. the IgG group. (d) TEER value was measured to evaluate the effects of MIR17HG and miR-153 (miR-377) on the permeability of glioma-conditioned normal BBB. (e) HRP flux was measured to evaluate the effects of MIR17HG and miR-153 (miR-377) on the permeability of glioma-conditioned normal BBB. (f and g) Western blot was performed to evaluate the effects of MIR17HG and miR-153 (miR-377) on tight junction-related proteins. Data of d-g represent the mean ± SD (n = 3, each). *P < 0.05 vs. the shMIR17HG-NC + pre-NC group, **P < 0.01 vs. the shMIR17HG-NC + pre-NC group