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. 2008 Jan 4;82(1):57–72. doi: 10.1016/j.ajhg.2007.09.012

Figure 3.

Figure 3

EMSA for Analyses of Protein/DNA Interactions and Relative Luciferase Activity of Intestinal Caco2 Cell Line Cotransfected with Lactase Promoter-Reporter Constructs Containing the Identified Variants

(A) Electrophoretical mobility shift assay (EMSA) for analyses of protein/DNA interactions of the identified variants (lanes 1–5) with double-stranded oligonucleotides primers and nuclear extracts from the intestinal cell line Caco2. The arrow indicates the Oct-1 complexes.

(B) The EMSA for the T/C−3712 showing the specific binding of HNF1α to both probes. The HNF1α supershift analyses were performed by adding a polyclonal antibody against rat HNF1α.

(C and D) Relative luciferase activity of intestinal Caco2 cell line cotransfected with lactase promoter-reporter constructs containing the 455 bp fragments of the regions flanking the T/G−13915 and 685 bp fragment of the region flanking T/C−3712.

(D) The G−13915-C−3712 and T−13915-T−3712 regions were analyzed together for the effect of overexpression of HNF1α and Oct-1 on the G−13915-C−3712 and T−13915-T−3712. The luciferase activity was corrected for transfection efficiency and normalized to the expression of pGL3-hLPH1085, n = 4.