sv40 enhancer sequence  (Addgene inc)

Journal: Nature Communications

Article Title: Auricular malformations are driven by copy number variations in a hierarchical enhancer cluster and a dominant enhancer recapitulates human pathogenesis

doi: 10.1038/s41467-025-59735-w

Figure Lengend Snippet: a Epigenetic profiling at the BCE core locus in early (D11) and late hCNCCs (P4). Previously reported mouse 594 bp ECR region resides in the mEC1 homologous sequence. b Schematic of hESCs differentiation to PA-like hCNCCs: The process involves transitioning H9 cell line hESCs to early hCNCCs using NCC induction (NCCI) medium, followed by BMP2/ChIR treatment to reach late hCNCCs stage, and finally achieving PA-like hCNCCs with retinoic acid (RA) introduction. c HMX1 expression analysis across four cell states, presented as mean ± S.E.M. ( n = 6). P -values were calculated using unpaired Student’s t -test (two-tailed). d, e Luciferase assays evaluating candidate enhancers at the BCE core locus in hESC ( d ), and PA-like hCNCC ( e ), including SV40 enhancer (positive control) and empty vector (negative control). Results from three independent experiments, each with four technical replicates ( n = 12), are shown. The box plot displays the median (center line), 25th-75th percentiles (box bounds), and whiskers extending to 1.5×interquartile range from each quartile (10th-90th percentiles shown as reference), with outliers plotted individually. f CRISPR/Cas9 strategy to delete two active enhancers (hEC1 and hEC2, ~9 kb), and gene expression comparison across four cell states in WT and knockout cell lines ( n = 3). P -values were calculated using unpaired Student’s t -test (two-tailed). The boxplot definition is same as d, e . Source data are provided as a Source Data file.

Article Snippet: The transfection mix included 0.5 μg of pGL3-enhancer plasmids with firefly luciferase gene sequences driven by an SV40 promoter (Promega, catalog no. E1761), 0.02 μg of pRL-CMV Renilla luciferase (Promega, catalog no. E2261) as an internal control, and 1.5 μL of FuGENE HD (Promega, catalog no. E2691), all diluted in Opti-MEM I Reduced-Serum Medium (Gibco, catalog no. 31985062).

Techniques: Sequencing, Expressing, Two Tailed Test, Luciferase, Positive Control, Plasmid Preparation, Negative Control, CRISPR, Gene Expression, Comparison, Knock-Out

Journal: The American Journal of Pathology

Article Title: Chitinase Inhibition Promotes Atherosclerosis in Hyperlipidemic Mice

doi: 10.1016/j.ajpath.2013.04.003

Figure Lengend Snippet: Chitinase inhibitor increases AP-1 and NF-κB transcriptional activity. A: RAW264.7 macrophages were treated with medium alone or 10 μmol/L allosamidin overnight. Nuclear protein was extracted and transcriptional activity of AP-1 and NF-κB were accessed by electromobility shift assay. B: RAW264.7 macrophages were transfected with AP-1 or NF-κB promoter–luciferase plasmid and 2 μg of β-galactosidase gene driven by SV40 promoter-enhancer sequence. The cells were then treated with medium alone or 10 μmol/L allosamidin overnight, and luciferase activity and β-galactosidase were measured. ∗P < 0.05 versus control.

Article Snippet: The cells were then transfected with 0.7 μg/well of AP-1 or NF-κB promoter–luciferase plasmid (Clontech) and 0.3 μg per well of β-galactosidase gene driven by SV40 promoter-enhancer sequence (Promega).

Techniques: Activity Assay, Electro Mobility Shift Assay, Transfection, Luciferase, Plasmid Preparation, Sequencing

Journal: The American Journal of Pathology

Article Title: Chitinase Inhibition Promotes Atherosclerosis in Hyperlipidemic Mice

doi: 10.1016/j.ajpath.2013.04.003

Figure Lengend Snippet: Chitinase inhibitor increases AP-1 and NF-κB transcriptional activity. A: RAW264.7 macrophages were treated with medium alone or 10 μmol/L allosamidin overnight. Nuclear protein was extracted and transcriptional activity of AP-1 and NF-κB were accessed by electromobility shift assay. B: RAW264.7 macrophages were transfected with AP-1 or NF-κB promoter–luciferase plasmid and 2 μg of β-galactosidase gene driven by SV40 promoter-enhancer sequence. The cells were then treated with medium alone or 10 μmol/L allosamidin overnight, and luciferase activity and β-galactosidase were measured. ∗P < 0.05 versus control.

Article Snippet: The cells were then transfected with 0.7 μg/well of AP-1 or NF-κB promoter–luciferase plasmid (Clontech) and 0.3 μg per well of β-galactosidase gene driven by SV40 promoter-enhancer sequence (Promega).

Techniques: Activity Assay, Electro Mobility Shift Assay, Transfection, Luciferase, Plasmid Preparation, Sequencing, Control