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Figure 2 | Molecular Cancer

Figure 2

From: The tumor suppressor gene KCTD11RENis regulated by Sp1 and methylation and its expression is reduced in tumors

Figure 2

Basal transcriptional activity of human KCTD11 promoter is depending on Sp1 TF. (A) Human KCTD11 promoter was identified by Promoter Inspector http://www.genomatix.de/. Strong matches for Sp1 transcription factor binding sites are boxed. The asterisk indicates the TSS. The CpG dinucleotides are grey highlighted and numbered. (B) Schematic representation of Luc reporter constructs driven by human full-length KCTD11 promoter or deletion mutants. Constructs were generated by PCR using genomic DNA from peripheral blood lymphocytes as template and specific primers [additional file 2] and subcloned in pGL3 basic vector. (C-D) Luciferase assays showing the transcriptional activity of full-length or deletion mutants of KCTD11 promoter. 293T HEK (human embryo kidney) cell line was cultured in DMEM supplemented with 10% FCS, 2 mM L-glutamine (Sigma), 100 U/ml penicillin and 100 μg/ml streptomycin (Sigma). 2 μg of KCTD11-Luc reporter constructs were transfected using Lipofectamine 2000 (Invitrogen). After 24 hrs, the cells were harvested and firefly luciferase activity was assayed using the Firelite Dual Luminescence Reporter Gene Assay System kit (Perkin-Elmer) normalized to Renilla luciferase activity. Each experiment has been done in triplicates. Values are the means ± S.D.

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