The cDNA encoding HOXA7 was subcloned into the expression vector pBabe and subcloning was confirmed with sequencing by Shanghai Genechem Co., Ltd. The pGL3 reporter construct plasmid (−3000/+100) consisted of a 3100-bp genomic DNA fragment of the Snail promoter (Promega, Madison, WI, USA). The pGL3-Snail promoter plasmid or its negative control pGL3-basic plasmid carrying the firefly luciferase reporter were co-transfected with an internal control, pRL-TK Renilla vector (Promega), by using Lipofectamine 2000 (Invitrogen). In addition, cells were respectively transfected with 600 ng of HOXA7 overexpression plasmid pBabe or its negative control pbabe-vector. Cell lysates were harvested 48 h after transfection. The firefly and renilla activities were measured by the Dual-Luciferase Reporter Assay System (Promega). Firefly luciferase activity was normalized to the Renilla luciferase activity. Each transfection was repeated three times.
Tang B., Qi G., Sun X., Tang F., Yuan S., Wang Z., Liang X., Li B., Yu S., Liu J., Huang Q., Wei Y., Zhai R., Lei B., Guo X, & He S. (2016). HOXA7 plays a critical role in metastasis of liver cancer associated with activation of Snail. Molecular Cancer, 15(1), 57.
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