Fig 1: Knockdown of CADM2 rescues the inhibitory effects of miR-25 downregulation on the proliferation and migration of glioma cells. T98G and U251MG cells were co-transfected with miR-25 and siCADM2 or siNC. Following transfection, the (A) mRNA and (B) protein expression levels of CADM2 were examined using reverse transcription-quantitative PCR and western blotting, respectively. The Cell Counting Kit-8 assay was used to determine cell proliferation in (C) T98G and (D) U251MG cells. Wound healing assays were used to study migration in (E) T98G and (F) U251MG cells. (G) Cell invasion was determined using Transwell assays. **P<0.01 vs. miR-25 in + siNC. CADM2, cell adhesion molecule 2; miR, microRNA; si, small interfering RNA; NC, negative control; in, inhibitor. Magnification for cell migration, x40. Magnification for cell invasion, x200.
Fig 2: miR-125a was increased in the cortex of ischemic rats. (A) The expression of miRNAs in cortex was measured using microarray analysis in control group (left) and model group (right). (B) Longa score in control group and model group. (C) The expression of miR-125a in cortex were measured using reverse transcription-quantitative polymerase chain reaction in the cortex, data were presented as the mean ± standard deviation, n=3 in each group. *P<0.05 vs. normal group. miR, microRNA; MCAO, middle cerebral artery occlusion; Cadm2, cell adhesion molecule 2.
Fig 3: WT1-AS-siRNA-reduced CADM2 expression in HTR-8/SVneo cells. HTR-8/SVneo cells were transfected with control siRNA, WT1-AS siRNA, WT1-AS siRNA + inhibitor control, or WT1-AS siRNA + miR-186-5p inhibitor. (a) Protein expression of CADM2 in HTR-8/SVneo cells was determined using western blotting. (b) mRNA expression of CADM2 in HTR-8/SVneo cells was determined using RT-qPCR. ** P < 0.01 vs control siRNA; ## P < 0.01 vs WT1-AS siRNA + inhibitor control. WT1-AS, WT1 antisense RNA; si, small interfering; miR, microRNA; RT-qPCR, reverse transcription-quantitative PCR.
Fig 4: Correlation between miR-186-5p and CADM2. (a) StarBase analysis predicted the relationship between miR-186a-5p and CADM2. (b) The association between miR-186-5p and CADM2 was verified via the dual-luciferase reporter assay. ** P < 0.01 vs mimic control. miR, microRNA; CADM2, cell adhesion molecule 2.
Fig 5: Cadm2 was decreased in the cortex of ischemic rats. (A) The expression of Cadm2 in cortex was measured by laser confocal microscopy (original magnification, ×200). (B) Longa score in control group and model group. The expression of Cadm2 was measured using a western blot assay in the cortex, and the immunoreactive bands were quantified with ImageJ software. By using western blot analysis, the expression of the Cadm2 protein was efficiently decreased in the model group, compared with the control group. *P<0.05 and **P<0.01 vs. control. MCAO, middle cerebral artery occlusion; Cadm2, cell adhesion molecule 2.
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