Fig. 2: TFEB binds human and mouse CYP7A1 promoter.
From: An FGF15/19-TFEB regulatory loop controls hepatic cholesterol and bile acid homeostasis
a The CLEAR sequence location relative to transcriptional start site of CYP7A1 is indicated in red. b, c EMSA assay of TFEB binding to CLEAR sequences in human and mouse CYP7A1 promoter. In-gel imaging of SYBR green signal is shown (b). rhTFEB recombinant human TFEB. NPC2 Nieman-Pick Disease Type-C2 (positive control). After SYBR green images were taken, protein in the same polyacrylamide gel was transferred to nitrocellulose membrane and immunoblotted with TFEB antibody (c). The arrow indicates the rhTFEB protein with the same mobility as the TFEB-probe complex shown in the left panel. This experiment was repeated once with similar results. d, e ChIP assay of TFEB binding to human and mouse proximal and distal promoter regions of the CYP7A1 gene chromatin in human and mouse livers. Pooled normal human livers (n = 3) and chow-fed mouse livers (n = 3) were used to isolate nuclei for ChIP assay as described in Methods. *, vs. IgG. p = 0.0006 for human proximal promoter; p = 0.003 for human distal promoter; p = 0.0035 for mouse proximal promoter; p = 0.002 for mouse distal promoter. Two-sided Student’s t-test was used. INPUT (0.1% total) was set as 1. Results were expressed as mean ± SD (technical triplicates). Source data for b–e are provided as a Source Data file.
