Fig 2: CACNA2D1 is a direct downstream target of miR-107. (a) Wild type and mutant of the 3'-UTR of CACNA2D1 mRNA and miR-107 binding site (site A: 202-209). (b) Luciferase activity controlled by the 3'-UTR of CACNA2D1 was inhibited by ectopic expression of miR-107 at site A. TU686 cells were co-transfected with wild-type or mutant CACNA2D1 3'-UTR and agomiR-107, antagomiR-107 and the corresponding negative controls. Renilla and firefly luciferase activities were measured using a dual-luciferase reporter assay system. The ratio of renilla luciferase activity to firefly luciferase activity was normalized, and the data are expressed as mean ± SEM (compared with the control group, *P < 0.05). (c) Wild type and mutant of the 3'-UTR of CACNA2D1 mRNA and miR-107 binding site (site B: 902-908). (d) Luciferase activity controlled by the 3'-UTR of CACNA2D1 was inhibited by ectopic expression of miR-107 at site B. TU686 cells were co-transfected with wild-type or mutant CACNA2D1 3'-UTR and agomiR-107, antagomiR-107, and the corresponding negative controls. The dual-luciferase reporter gene system indicates that miR-107 can bind to the site B of CACNA2D1 3'-UTR and inhibit the luciferase activity (compared with the control group, *P < 0.05). miR-107, microRNA-107

Fig 3: CACNA2D1 and miR-107 are inversely correlated in LSCC cell lines. (a and b) Transfection efficiency among the following five groups: TU212 and TU686 cells transfected with blank control, agomiR NC, agomiR-107, antagomiR NC, or antagomiR-107, and the expression of miR-107 was significantly increased or decreased, respectively (compared with the control group *P < 0.05). (c and d) qPCR analysis of relative expression level of CACNA2D1 mRNA after transfection of TU212 and TU686 cells (compared with the control group *P < 0.05). (e and f) Western blot analysis of relative expression of α2δ1 protein in transfected TU212 and TU686 cells (compared with the control group, *P < 0.05). LSCC, laryngeal squamous cell carcinoma; miR-107, microRNA-107; NC, negative control.