notch1 shrna  (Santa Cruz Biotechnology)

Journal: iScience

Article Title: Revealing the role of cancer-associated fibroblast senescence in prognosis and immune landscape in pancreatic cancer

doi: 10.1016/j.isci.2024.111612

Figure Lengend Snippet:

Article Snippet: Viral particles expressing Notch1-shRNA and Notch1 were purchased from Genechem Corporation (Shanghai, China) and used to transduce PANC-1 and BxPC-3 according to the manufacturer’s protocol.

Techniques: Purification, Blocking Assay, Virus, Plasmid Preparation, Recombinant, Saline, Staining, Marker, Bicinchoninic Acid Protein Assay, CCK-8 Assay, Infection, Enzyme-linked Immunosorbent Assay, Gene Expression, Microarray, Expressing, Software

Journal: PLoS ONE

Article Title: Directed Differentiation of Patient-Specific Induced Pluripotent Stem Cells Identifies the Transcriptional Repression and Epigenetic Modification of NKX2-5, HAND1, and NOTCH1 in Hypoplastic Left Heart Syndrome

doi: 10.1371/journal.pone.0102796

Figure Lengend Snippet: Transcriptional activation of SRE promoter luciferase construct by combinatorial transfection of NKX2-5, HAND1, and NOTCH1 in HLHS- and BV-derived CPCs (A) or iPS cells (B). Co-transfection of TNNT2 luciferase reporter with NKX2-5, HAND1, and NOTCH1 in CPCs (C) or iPS cells (D) is shown. NPPA luciferase construct was co-transfected with NKX2-5, HAND1, and NOTCH1 alone or in combination into CPCs (E) or iPS cells (F). (G-I) BV-derived CPCs were transfected with either control or shRNAs specific to inhibit NKX2-5, HAND1, and NOTCH1 expression. Results were normalized using an internal control (SEAP or hRluc) and obtained from more than triplicate sets of experiments. *, p<0.05 vs. the same HLHS sample without transfection of the gene of interest. †, p<0.05 vs. BV sample transfected with control vector alone. ‡, p<0.05 vs. both HLHS samples with the same treatment. §, p<0.01 vs. BV sample transfected with control vector alone.

Article Snippet: Human cells were transfected with human NKX2-5 cDNA (SC122678), human NKX2-5 shRNA (short hairpin RNA; TR311165B), human HAND1 cDNA (SC122690), human HAND1 shRNA (TR316857C), human NOTCH1 cDNA (SC308883), or human NOTCH1 shRNA (TR302916D) along with single- or dual-reporter constructs (all from OriGene Technologies, Inc.).

Techniques: Activation Assay, Luciferase, Construct, Transfection, Derivative Assay, Cotransfection, Expressing, Plasmid Preparation

Journal: PLoS ONE

Article Title: Directed Differentiation of Patient-Specific Induced Pluripotent Stem Cells Identifies the Transcriptional Repression and Epigenetic Modification of NKX2-5, HAND1, and NOTCH1 in Hypoplastic Left Heart Syndrome

doi: 10.1371/journal.pone.0102796

Figure Lengend Snippet: (A) Core transcriptional factors expressed in cardiac progenitor cells serve as targets in response to inductive signals to initiate cardiogenesis. NKX2-5 is predominantly expressed in the primary heart field and controls progenitor cell proliferation. Genes regulate atrioventricular (AV) canal and valve development. Reduced expression may contribute to mitral- and aortic-valve stenosis/atresia often seen in HLHS. NOCTH modulates left heart outflow tract development and the resultant obstruction may cause secondary ventricle hypoplasia. HAND1/2 specify left and right ventricular chamber morphogenesis, and the absence of these genes may lead to a hypoplastic ventricle. (B–D) Schematic diagrams of SRE , TNNT2 , and NPPA transcriptional activation. HLHS-derived CPCs and iPS cells showed significantly reduced luciferase activities compared with BV-derived cells. Co-transfection analysis of reporter constructs with NKX2-5, HAND1, and NOTCH1, proposed core transcriptional factors, could synergistically restore the transcriptional activation in these reporters equivalent to the levels in BV-derived cells. (E) Major chromatin features in differentiated HLHS- and BV-derived iPS cells are shown. Upon cardiomyocyte differentiation, HLHS-derived iPS cells failed to enrich the active histone marks such as H3K4me2 and acH3, whereas repressive histone marks such as H3K27me3 increased, resulting in compact chromatin that lost enhancer marks and gained repressor marks on the NKX2-5 promoter.

Article Snippet: Human cells were transfected with human NKX2-5 cDNA (SC122678), human NKX2-5 shRNA (short hairpin RNA; TR311165B), human HAND1 cDNA (SC122690), human HAND1 shRNA (TR316857C), human NOTCH1 cDNA (SC308883), or human NOTCH1 shRNA (TR302916D) along with single- or dual-reporter constructs (all from OriGene Technologies, Inc.).

Techniques: Expressing, Activation Assay, Derivative Assay, Luciferase, Cotransfection, Construct