Figure 3.
HBB mutation binds with TBP and verifies the competitive interaction between HBB, HBG, and LCR. (A) pGL3-basic plasmid was inserted in WT sequences, and HBB was c.-78A>G mutant HBB promoters cotransfected them with the TK (Renilla Luciferase-Thymidine Kinase Plasmid) reference plasmid into 293T. The results reveal the changes in luminescence optical density‌ values of the WT (HBB WT promoter), -28(HBB: c.-78A>G), and -29(HBB:c.79A>G) mutation to determine the changes in promoter activity. (B) Co-transfection of pGL3-basic negative control, HBB WT, HBB-28(HBB:c.-78A>G), or HBB-29(HBB:c.-79A>G) and plasmids with pcDNA3.1 NC or TBP plasmid into 293T cells. (C) ChIP-qPCR assay for TBP or IgG occupancy at HBB:c.-78A>G and mock-edited control HUDEP-2 cells. (D) 4C profiles of HS3 in populations of HBB:c.-78A>G–edited and mock-edited control HUDEP-2 cells (n = 2). The positions of the β-globin locus genes are indicated below the profiles. The 4C track file comes from the.wig.gz file of the data analysis results of the Pipe 4C pipeline. Fold change = (experimental group 4C track/control group 4C track) – 1. When the fold change is >0, it indicates an upregulation of interaction strength in the experimental group, whereas downregulation of interaction strength in the experimental group is illustrated if the fold change is <0. (E) Quantitative analysis of relative interaction frequency between the captured bait and the interacting regions. Data shown are the means ± SEM. Statistical significance was calculated using the Student t test. Asterisks indicate levels of statistical significance. ∗∗P < .01; and ∗∗∗∗P < .0001. IgG, immunoglobulin G; ns, no significance.

HBB mutation binds with TBP and verifies the competitive interaction between HBB, HBG, and LCR. (A) pGL3-basic plasmid was inserted in WT sequences, and HBB was c.-78A>G mutant HBB promoters cotransfected them with the TK (Renilla Luciferase-Thymidine Kinase Plasmid) reference plasmid into 293T. The results reveal the changes in luminescence optical density‌ values of the WT (HBB WT promoter), -28(HBB: c.-78A>G), and -29(HBB:c.79A>G) mutation to determine the changes in promoter activity. (B) Co-transfection of pGL3-basic negative control, HBB WT, HBB-28(HBB:c.-78A>G), or HBB-29(HBB:c.-79A>G) and plasmids with pcDNA3.1 NC or TBP plasmid into 293T cells. (C) ChIP-qPCR assay for TBP or IgG occupancy at HBB:c.-78A>G and mock-edited control HUDEP-2 cells. (D) 4C profiles of HS3 in populations of HBB:c.-78A>G–edited and mock-edited control HUDEP-2 cells (n = 2). The positions of the β-globin locus genes are indicated below the profiles. The 4C track file comes from the.wig.gz file of the data analysis results of the Pipe 4C pipeline. Fold change = (experimental group 4C track/control group 4C track) – 1. When the fold change is >0, it indicates an upregulation of interaction strength in the experimental group, whereas downregulation of interaction strength in the experimental group is illustrated if the fold change is <0. (E) Quantitative analysis of relative interaction frequency between the captured bait and the interacting regions. Data shown are the means ± SEM. Statistical significance was calculated using the Student t test. Asterisks indicate levels of statistical significance. ∗∗P < .01; and ∗∗∗∗P < .0001. IgG, immunoglobulin G; ns, no significance.

or Create an Account

Close Modal
Close Modal