Validation of TSKU as direct target of miR-2110.

(A) The predicted interactions between miR-2110 and its three target sites in the 3’UTR of TSKU mRNA. The seed sequences are underlined. pos, position. (B) The schematic diagram showing the mutations on predicted target sites of TSKU 3’UTR in the corresponding luciferase reporters. (C) Validation of the target sites by luciferase assay. BE(2)-C cells were co-transfected with the indicated vectors and oligos (miR-2110 mimic or control oligo). Three days after transfection, cells were lysed and luciferase activity was measured and analyzed for the indicated comparison. *, p p D-E) Effect of TSKU 3’UTR over-expression on the function of miR-2110 mimic in inducing neurite outgrowth and in reducing cell viability. BE(2)-C cells were co-transfected with vectors (TSKU-WT or the control vector without 3’UTR of TSKU) and oligos (miR-2110 mimic or control oligo). Neurite outgrowth and cell viability were measured as above. D, Neurite outgrowth under each treatment. *, p E, Cell viability under each treatment. *, p p < 0.05 comparing the indicated two groups.