The promoter sequence of Slug gene (from -2000 to -1) was obtained from the UCSC website. Partial sequences of the promoter were amplified by PCR, and the fragments were cloned into the pGL3-basic vector. MDA-MB-231 cells were co-transfected with pGL3-basic or pGL3-Slug-Luc together with sh-HES1 or sh-NC, and pRL-TK plasmid. After 24 h, the cells were lysed and luciferase activities were analyzed using a Promega dual-luciferase assay kit according to the manufacturer's instructions.

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