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. 2015 Dec 11;5:17972. doi: 10.1038/srep17972

Figure 2. The T/C mutation in the DKK2 5′ flanking sequence DKK2 c.−1130 T > C affected promoter activity in CHO and HeLa cells.

Figure 2

(A) Genotyping for DKK2 c.−1130 T > C using PCR-TaqI-RFLP. Genotype TC: 1225 bp+759 bp+466 bp; Genotype TT: 1225 bp; Genotype CC: 759 bp+466 bp. M: DNA molecular marker DL 2,000. (B) Schematic structure of the DKK2 c.−1130 T > C mutation in the fourth C/EBPβ binding site. (C) pGL3-D3-T and pGL3-D3-C were transfected into CHO and HeLa cells. Compared to pGL3-D3-C, pGL3-D3-T transfections resulted in significantly higher relative luciferase activity (P < 0.01). (D) pGL3-D3-T and pGL3-D3-C were co-transfected with the pGL3-C/EBPβ expression plasmid, resulting in increased DKK2 promoter luciferase activity. In the pc-C/EBPβ groups, the T allele group showed a significantly higher level of relative luciferase activity than the C allele group in CHO cells (P < 0.01). (E) siRNA (2 μl) was co-transfected with 0.2 μg pGL3-D3-T or pGL3-D3-C for 24 h in CHO and HeLa cells, and DKK2 promoter luciferase activity was inhibited. The pGL3-basic plasmid was used as a negative control. The pRT-TK plasmid served as an inner control during analysis of transfection efficiency. **P < 0.01.

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