Fig. 2

miR-218-5p has no effect on the proliferation of ESCs but can inhibit the migration of ESCs. A Western blot analysis and quantification of LASP1 protein level in lysates of different ESCs. B Western blot analysis and quantification of LASP1 protein level in lysates of Q-Z1 with or without 50 nM miR-218-5p mimic. C Western blot analysis and quantification of LASP1 protein level in lysates of ThESc with or without 50 nM miR-218-5p inhibitor. GAPDH was used as the loading control. P-values were determined by Student’s t-test, *p < 0.05, ***p < 0.001. D Predicted miR-218-5p target recognition sites in the 3’-UTR of human LASP1. Mutation in the miR-218-5p target site is red. 3’UTR of LASP1 luciferase constructs, as indicated, were transfected into 293 T cells together with indicated plasmid for 48 h and subjected to a luciferase reporter assay. The results were normalized to the Renilla luciferase activity and are expressed as the fold change in relative luciferase activity compared with the control. E ThE cells were transfected with control inhibitor or miR-218-5p inhibitor. F Q-Z1 cells were transfected with control mimic or miR-218-5p mimic. After 24 h of transfection, cells were starved for 24 h before cell migration assay was performed without matrigel transwell filters, Scale bar = 200 μm. The migrated cells were stained and counted. Quantification was done in and is shown with counting six nonoverlaping fields. G and H EdU incorporation assays of ThE or Q-Z1 cells were transiently transfected with control inhibitor/miR-218-5p inhibitor or control mimic/miR-218-5p mimic, DAPI staining was included to visualize the cell nucleus (Blue), Scale bar = 100 μm. Each bar indicates mean ± SEM. of a representative experiment performed in triplicate. P-values were determined by Student’s t-test. *** p < 0.001