Fig 1: TCF7L2 is a transcriptional factor of miR-203(A) The protein expression level of TCF7L2 in the ileum of control and obese rats was analyzed by Western blot, n = 5.(B and C) HIECs were transfected with TCF7L2 overexpression plasmid. The levels of (B) TCF7L2 mRNA and (C) miR-203 in HIECs in different groups were determined by qRT-PCR, n = 3.(D and E) HIECs were transfected dODN. The levels of (D) miR-203, n = 5 and (E) luciferase activities in HIECs in different groups were detected, n = 3.(F) EMSA to test in vitro binding of TCF7L2 to the promoter region of human host genes for miR-203. Note shifted band representing protein-DNA binding (yellow arrow), abolished with the excess nonlabeled probe. Supershifted band (red arrow) represents TCF7L2-miR-203 binding in presence of the anti-TCF7L2 antibody.(G) ChIP assay to test the binding of TCF7L2 to the human host gene of miR-203. Top: PCR products of TCF7L2-miR-203 binding sites following immunoprecipitation with anti- TCF7L2 antibody. Bottom: host gene binding measured by qPCR following ChIP, expressed as fold-changes over IgG-negative control. Input: genomic DNA without immunoprecipitation, n = 3. The data are expressed as mean ± SEM, *p < 0.05, **p < 0.01, ***p < 0.001 vs control in (A-E), vs Input in (G) by one-way ANOVA with Turkey’s multiple comparison test.