Transcriptional regulation of human miR-200b-a-429 and miR-200c-141 by Ascl2.
A, schematic representation of the human miR-200b-a-429 promoter constructs that were transfected into shRNA-Ascl2/LS174T cells or shRNA-Ctr/LS174T cells in this study. Four promoter deletion-luciferase constructs were generated to identify the sites of transcriptional regulation within the human miR-200b-a-429 promoter that respond to Ascl2 knockdown (A). B, schematic representation of the human miR-200c-141 promoter construct (WT-Luc) and the p53-response element mutant construct (Mut-Luc) transfected into shRNA-Ascl2/LS174T cells or shRNA-Ctr/LS174T cells to identify the sites of transcriptional regulation within the human miR-200c-141 promoter that respond to Ascl2 knockdown. The cells were harvested after 48 h, and the relative luciferase activity was determined using the Dual-Luciferase reporter assay system with a single sample luminometer. The data represent the mean ± S.E. of three independent experiments (*, p < 0.05; **, p < 0.01). Error bars represent S.D.