Fig. 5

SNAI2/miR-4306 axis negative feedback regulates TCF7 expression. A The expression of miRNAs (miR-4306, miR-641 and miR-2278) was evaluated by RT-qPCR in HLF cells with SNAI2 knockdown or overexpression. B JASPAR predicted the DNA binding motif of SNAI2 and the possible binding sites of SNAI2 in miR-4306 promoter. C ChIP assays validated the interaction between miR-4306 promoter and SNAI2. D Luciferase reporter assays confirmed that SNAI2 bound to miR-4306 promoter at the putative sites. E RT-qPCR used to detect the miR-4306 expression in HLF cells transfected with vector, shTCF7, shTCF7 + SNAI2, TCF7, and TCF7 + shSNAI2. F The expression of miR-4306 and TCF7 was evaluated by RT-qPCR in HLF cells with miR-4306 knockdown or overexpression. G The protein expression of TCF7 was evaluated by Western Blot in HLF cells with miR-4306 knockdown or overexpression. H RIP assays demonstrated the interaction between miR-4306 and TCF7. I Predicted miR-4306 target sequence in TCF7-3′ UTR (TCF7-WT) and mutant containing mutated nucleotides in the seed sequence of TCF7-3′ UTR (TCF7-Mut), and the activity of the TCF7-3′ UTR-WT and TCF7-3′ UTR-Mut luciferase reporter gene in indicated cells. shSNAI2, SNAI2 knockdown adenovirus; shNC, corresponding negative control of SNAI2 knockdown adenovirus; SNAI2, SNAI2 overexpressed adenovirus; mock, corresponding negative control of SNAI2 overexpressed adenovirus; shTCF7, TCF7 knockdown adenovirus; TCF7 overexpressed adenovirus; NC inhi., negative control inhibitor; miR inhi., miR-4306 inhibitor; NC mim., negative control mimics; miR mim., miR-4306 mimics; ^#P < 0.05, ^##P < 0.01, and ^###P < 0.001