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).