Research Paper Volume 12, Issue 24 pp 25547—25563

Figure 3. Characterization of the regulatory relationship between MAGI2-AS3, mir-31-5p and TNS1 in BCa cells. (A–C) QRT-PCR analysis shows the expression levels of MAGI2-AS3, miR-31-5p and TNS1 mRNA in control (si-NC-transfected), MAGI2-AS3 knockdown (siMAGI2-AS3-transfected) and MAGI2-AS3-overexpressing (oeMAGI2-AS3) T24 and J82 cell lines. (D, E) QRT-PCR analysis shows miR-31-5p and TNS1 mRNA levels in control, MAGI2-AS3-silenced, and MAGI2-AS3-silenced plus miR-31-5p-silenced T24 and J82 cell lines. (F) Luciferase reporter assay results show the relative luciferase activity in T24 cells transfected with vectors containing wild-type MAGI2-AS3 and control or miR-31-5p compared to those transfected with vectors containing mutant MAGI2-AS3 and control or miR-31-5p. (G) Luciferase reporter assay results show the relative luciferase activity in T24 cells transfected with vectors containing wild-type TNS1 and control or miR-31-5p compared to those transfected with mutated TNS1 and control or miR-31-5p. (H, I) RNA pull-down assay results show the relative levels of MAGI2-AS3 and TNS1 pulled down by NC-miR-31-5p, Bio-miR-31-5p-WT or Bio-miR-31-5p-Mut. (J) Representative western blot shows TNS1 protein levels in control, MAGI2-AS3 knockdown (siMAGI2-AS3-transfected), MAGI2-AS3-overexpressing (oeMAGI2-AS3), and MAGI2-AS3-overexpressing plus miR-31-5p-overexpressing (oeMAGI2-AS3 plus oemiR-31-5p) T24 and J82 cells. *p<0.05. **p<0.01. GAPDH was used as internal control.