Fig. 11

Identification of ALG3 as a novel target of miR-142-5p. A microRNA-seq was performed to identify ALG3-associated miRNAs following ALG3 knockdown. B Venn diagram analysis intersecting up-regulated miRNAs from microRNA-seq (n = 64) with TargetScan-predicted ALG3-targeting miRNAs (n = 11). C Predicted binding sites between miR-142-5p and ALG3 3’-UTR from TargetScan. D Validation of miR-142-5p downregulation in bladder cancer from GSE211692 data set. E, F CCK8 assays were utlized to evaluate the effects of overexpression of miR-142-5p (E) and downregulation of miR-142-5p (F) on bladder cancer cell proliferation. G, H Colony formation assays were performed to assess the impacts of miR-142-5p mimics (G) and miR-142-5p inhibitor (H) on the colony forming abilities of bladder cancer cells. I, J Transwell migration and invasion assays were conducted to estimate the effects of overexpression and downregulation of miR-142-5p on cell metastatic capabilities. K Dual-gene luciferase activity assay was utilized to evaluate the direct binding between miR-142-5p and ALG3 3’-UTR. L, M qRT-PCR was conducted to assess the up-regulate effect of miR-142-5p mimics (L) and down-regulate effect of miR-142-5p inhibitor (M) on miR-142-5p expression. N–Q qRT-PCR (N, O) and western blot (P, Q) was conducted to evaluate the ALG3 expression after miR-142-5p mimics (N, P) and miR-142-5p inhibitor (O, Q) transfection for 48 h. **P < 0.01, ***P < 0.001