Fig. 5

SP1 induces the expression of miR-4310 by binding to its promoter region. a Schematic diagram of the promoter regions of miR-4310 with the putative SP1 TFBSs (site A and site B) and the structure of the wild-type (WT) and TFBS mutant (mut A, mut B, and mut A + B) luciferase reporters driven by the promoter. b Expression of SP1 was detected by qPCR assays. c, d The expression of PI3K/AKT pathway and EMT related proteins were examined by Western blot. GAPDH were used as a loading control. e ChIP assay along with PCR and agarose gel electrophoresis showed amplification of SP1-binding sites A and B. f Protein-DNA interactions between SP1 and the miR-4310 promoter were determined by electrophoretic mobility shift assays (EMSA). g Luciferase reporter assay was used to determine the binding of SP1 to miR-4310 promoter region. Statistical methods: One-way ANOVA and Dunnett’s multiple comparison test. Data were presented as mean ± s.d. NS no statistical significance, *p<0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001