2.13 Luciferase reporter assay
The mutant-type (MuT) 3′-UTR of EZH2, CTGF and Smad4 was constructed. The WT or MuT 3′-UTR of EZH2, CTGF and Smad4 were inserted into pMIRGLO vectors (Promega, USA). The miR-26a mimic or miR-negative control and WT or MuT 3′-UTR of EZH2, CTGF and Smad4 vector were co-transfected into HEK293T cells (Wanleibio, China). After 48 h, cells were lysed, and luciferase activity was determined by using a luciferase assay kit (Promega, USA) and multi-function microplate reader (TECAN, M200Pro, Switzerland).