Figure 1
Figure 1. Activity, structure, and function of BCL2 promoter. (A) Structure of the BCL2 promoter and position of constructs for determination of transcriptional activity. Shown are the regions comprising the “stimulatory” P1 promoter and the “inhibitory” P2 promoter and the position of the (−938)C>A polymorphism. ATG is the translation initiation codon; the arrow indicates the transcription start site. The reporter construct, cloned into the pSEAP-basic vector and the pGL3-basic vector, is shown encompassing nt −690 to −1001 (ie, part of the P2 promoter). See the text for further details. (B) Genotype-dependent activity of BCL2 5′ regulatory region reporter constructs expressed in HEK293 cells. Constructs were cloned into the pSEAP-basic vector and reporter activity was quantified by measuring the concentration of secreted alkaline phosphatase in the cell culture medium (details in “Patients, materials, and methods”). In each experiment, cells were transfected in parallel with constructs harboring the A allele or the C allele. A total of 9 independent pairs of transfection experiments were conducted. The reporter activity was normalized to pSEAP-basic activity. Each symbol represents reporter activity of 1 experiment. Connecting lines indicate results from transfections performed in parallel. The P value was calculated using the Wilcoxon matched pairs test. While absolute reporter activity was variable in different experiments, reporter activity of C-allele constructs was always higher than that of A-allele constructs. (C) Genotype-dependent activity of BCL2 5′ regulatory region reporter constructs expressed in Karpas 422 cells. Constructs were cloned into the pGL3 vector and reporter activity was quantified by measuring luciferase activity (see “Patients, materials, and methods”). Reporter activities were normalized to pGL3-basic activity. In each experiment, cells were transfected in parallel with constructs harboring the A allele or the C allele. A total of 6 independent pairs of transfection experiments were conducted. Each symbol represents reporter activity of 1 experiment. Connecting lines indicate results from transfections performed in parallel. In general, reporter activity was lower than that following expression of constructs in HEK293 cells (see panel B). The P value was calculated using the Wilcoxon matched pairs test. While absolute reporter activity was variable in different experiments, reporter activity of C-allele constructs was always higher than that of A-allele constructs.

Activity, structure, and function of BCL2 promoter. (A) Structure of the BCL2 promoter and position of constructs for determination of transcriptional activity. Shown are the regions comprising the “stimulatory” P1 promoter and the “inhibitory” P2 promoter and the position of the (−938)C>A polymorphism. ATG is the translation initiation codon; the arrow indicates the transcription start site. The reporter construct, cloned into the pSEAP-basic vector and the pGL3-basic vector, is shown encompassing nt −690 to −1001 (ie, part of the P2 promoter). See the text for further details. (B) Genotype-dependent activity of BCL2 5′ regulatory region reporter constructs expressed in HEK293 cells. Constructs were cloned into the pSEAP-basic vector and reporter activity was quantified by measuring the concentration of secreted alkaline phosphatase in the cell culture medium (details in “Patients, materials, and methods”). In each experiment, cells were transfected in parallel with constructs harboring the A allele or the C allele. A total of 9 independent pairs of transfection experiments were conducted. The reporter activity was normalized to pSEAP-basic activity. Each symbol represents reporter activity of 1 experiment. Connecting lines indicate results from transfections performed in parallel. The P value was calculated using the Wilcoxon matched pairs test. While absolute reporter activity was variable in different experiments, reporter activity of C-allele constructs was always higher than that of A-allele constructs. (C) Genotype-dependent activity of BCL2 5′ regulatory region reporter constructs expressed in Karpas 422 cells. Constructs were cloned into the pGL3 vector and reporter activity was quantified by measuring luciferase activity (see “Patients, materials, and methods”). Reporter activities were normalized to pGL3-basic activity. In each experiment, cells were transfected in parallel with constructs harboring the A allele or the C allele. A total of 6 independent pairs of transfection experiments were conducted. Each symbol represents reporter activity of 1 experiment. Connecting lines indicate results from transfections performed in parallel. In general, reporter activity was lower than that following expression of constructs in HEK293 cells (see panel B). The P value was calculated using the Wilcoxon matched pairs test. While absolute reporter activity was variable in different experiments, reporter activity of C-allele constructs was always higher than that of A-allele constructs.

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