Fig. 4

The miR-15a-PAI-2 axis promotes migration of CCA cells. a Luciferase assay of pmirGLO vector containing wild type 3’-UTR of PAI-2 and its deletion mutant of putative miR-15a binding sequences in mature miR-15a-transfected cells. The data were normalized to scrambled miRNA transfected cells. Bars represent mean ± SD of five measurements. b Secreted PAI-2 in CM from miR-15a mimic-transfected C096 CCFs examined by Western blot analysis. CMs from cells with no transfection and scrambled miRNA-transfected cells were used as a control. Ponceau S staining of each sample on the membrane was shown. Densitometric analysis of PAI-2 normalized against the total protein loading is shown. c The effect of rPAI-2 on migration of KKU-213 CCA cells was examined by a wound healing assay at different time points. Graphs show % of wound areas in KKU-213 cells with or without rPAI-2 in. Bars represent mean ± SD of three measurements. d The effect rPAI-2 on CCA cells in the chamber migration assay. Graphs show the number of migrated cells with or without rPAI-2 treatment at 7 h. Bars represent mean ± SD of three measurements. e The effect of CM from miR-15a mimic or scrambled miRNA transfected C096 CCFs on the migration of CCA cells with or without rPAI-2. The migration was evaluated at 12 h by a wound healing assay as previously described. Bars represent mean ± SD of three measurements. *P < 0.05