Research Paper Advance Articles

Figure 4. RNA-seq and ChIP-seq were performed to screen the downstream target genes of EGR1. (A) Heatmap analysis was performed using DeepTools software to compare the signal intensity adjacent to the transcription start site between the EGR1 overexpression and control groups. (B) Venn diagram showing screened candidate genes identified by the intersection of Chip-seq and RNA-seq data. (C) Protein-protein interaction analysis indicating interactions between candidate genes and EGR1. (D) Potential binding sites for EGR1 on the nuclear receptor 4A3 (NR4A3) promoter were identified using the JASPAR database to analyze the ChIP-seq data. (E) Schematic diagram of three potential EGR1 binding sites (site 1, site 2, and site 3) in the NR4A3 promoter. (F) ChIP-qPCR was utilized to assess the binding of EGR1 to the NR4A3 promoter region. (G) The activation effect of EGR1 on the NR4A3 promoter was evaluated using a dual-luciferase reporter assay. (H) NPCs were transfected with plasmids containing either the full-length or site 1-mutated NR4A3 promoter to measure the luciferase activity of the NR4A3 promoter upon EGR1 overexpression. The data are expressed as the mean ± SD (n = 3). ^**p < 0.01, ^****p < 0.0001.