Figure 4.

Target prediction and experimental validation. (a) Comparison of the common genes downregulated by miR-296-5p, miR-324-3p, and miR-3928-3p in PC10 or ACC-LC-73 cells. (b) Barplot demonstrating the ability of miRNA mimics to reduce FAM46C mRNA expression in PC10 or ACC-LC-73 cells. (c) Boxplot showing the decrease in FAM46C expression in Sq compared to Ad and normal tissues (N). P-values were calculated with the Wilcoxon rank sum test: (**) P-value < 0.01. (d) Barplot showing the ability of each miRNA to bind to AGO2 protein. (e) Barplot showing the ability of FAM46C mRNA to bind to AGO2 protein via miRNA. (f) Barplot demonstrating the ability of FAM46C siRNA to inhibit FAM46C mRNA expression in PC10 cells (left). Barplot demonstrating the ability of FAM46C siRNA to enhance MYC mRNA expression in PC10 cells (right). Error bars show standard deviations. (g) Western blot showing the decrease in FAM46C protein (f: full-length protein, s: short-length protein, ref.¹¹) expression and the increase in MYC protein expression upon transfection with FAM46C siRNA. (h) Polygonal line demonstrating the ability of the combination of the three miRNA inhibitors to suppress the proliferation of PC10 cells. Western blot showing the decrease in MYC protein expression upon transfection of the combination of the three miRNA inhibitors miRNAs in PC10 cells (see also Supplementary Fig. S8).