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    Protocols in Current Issue
    Lentiviral Knockdown of Transcription Factor STAT1 in Peromyscus leucopus to Assess Its Role in the Restriction of Tick-borne Flaviviruses
    Authors:  Adaeze O. Izuogu and R. Travis Taylor, date: 12/05/2017, view: 100, Q&A: 0
    Cellular infection with tick-borne flaviviruses (TBFVs) results in activation of the interferon (IFN) signaling pathway and subsequent upregulation of numerous genes termed IFN stimulated genes (ISGs) (Schoggins et al., 2011). Many ISGs ...
    In vivo Leukemogenesis Model Using Retrovirus Transduction
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 98, Q&A: 0
    Various genetic alterations such as chromosomal translocation cause leukemia. For examples, gene rearrangements of the mixed-lineage leukemia (MLL) gene generate MLL fusion genes, whose products are potent oncogenic drivers in ...
    Myeloid Progenitor Transformation Assay
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 116, Q&A: 0
    Numerous oncogenes have been identified to cause leukemia. For example, chromosomal translocation generates various fusion genes of the mixed-lineage leukemia (MLL) gene and a partner gene in leukemia, whose gene products transform primary ...
    Measuring the Endocytic Recycling of Amyloid Precursor Protein (APP) in Neuro2a Cells
    Authors:  Florent Ubelmann, Tatiana Burrinha and Claudia Guimas Almeida, date: 12/05/2017, view: 135, Q&A: 0
    The established primary trigger of Alzheimer’s disease’s is β-amyloid (Aβ) (Mucke and Selkoe, 2012). Amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at ...
    A Novel Protocol to Quantitatively Measure the Endocytic Trafficking of Amyloid Precursor Protein (APP) in Polarized Primary Neurons with Sub-cellular Resolution
    Authors:  Florent Ubelmann, Tatiana Burrinha and Cláudia Guimas Almeida, date: 12/05/2017, view: 86, Q&A: 0
    Alzheimer’s disease’s established primary trigger is β-amyloid (Aβ) (Mucke and Selkoe, 2012). The amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at endosomes ...
    Notch Ligand Binding Assay Using Flow Cytometry
    Authors:  Shweta Varshney and Pamela Stanley, date: 12/05/2017, view: 156, Q&A: 0
    Notch signaling is an evolutionarily conserved signaling pathway that plays an indispensable role during development, and in the maintenance of homeostatic processes, in a wide variety of tissues (Kopan, 2012; Hori et al., 2013). The ...
    Formalin Murine Model of Pain
    Pain research is mostly based on experimental assays that use animal models, which may allow deciphering the physiopathology of this condition and to propel drug discovery. The formalin nociception test is considered one of the most predictive ...
    GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells
    Human induced pluripotent stem cells (hiPSCs) are a promising tool in cell-based therapies for degenerative diseases. A safe application of hiPSCs in vivo, requires the detection of the presence of residual undifferentiated pluripotent ...
    Mandibular Explant Assay for Investigating Extrinsic Stimuli on Bone and Cartilage Development
    Authors:  Jamie Jiang, Jessica W. Bertol and Walid D. Fakhouri, date: 12/05/2017, view: 139, Q&A: 0
    A major issue in developmental biology is to determine how embryonic tissues respond to molecular signals in a timely manner and given the position-restricted instructions during morphogenesis, of which Meckel’s cartilage is a classical example. The ...
    Lentiviral Knockdown of Transcription Factor STAT1 in Peromyscus leucopus to Assess Its Role in the Restriction of Tick-borne Flaviviruses
    Authors:  Adaeze O. Izuogu and R. Travis Taylor, date: 12/05/2017, view: 100, Q&A: 0
    [Abstract] Cellular infection with tick-borne flaviviruses (TBFVs) results in activation of the interferon (IFN) signaling pathway and subsequent upregulation of numerous genes termed IFN stimulated genes (ISGs) (Schoggins et al., 2011). Many ISGs function to prevent virus pathogenesis by acting in a broad or specific manner through protein-protein ...
    In vivo Leukemogenesis Model Using Retrovirus Transduction
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 98, Q&A: 0
    [Abstract] Various genetic alterations such as chromosomal translocation cause leukemia. For examples, gene rearrangements of the mixed-lineage leukemia (MLL) gene generate MLL fusion genes, whose products are potent oncogenic drivers in acute leukemia. To better understand the mechanism of disease onset, several murine leukemia models ...
    Myeloid Progenitor Transformation Assay
    Authors:  Hiroshi Okuda and Akihiko Yokoyama, date: 12/05/2017, view: 116, Q&A: 0
    [Abstract] Numerous oncogenes have been identified to cause leukemia. For example, chromosomal translocation generates various fusion genes of the mixed-lineage leukemia (MLL) gene and a partner gene in leukemia, whose gene products transform primary myeloid progenitors into an immortalized state. To characterize the transforming ability of leukemic ...
    Measuring the Endocytic Recycling of Amyloid Precursor Protein (APP) in Neuro2a Cells
    Authors:  Florent Ubelmann, Tatiana Burrinha and Claudia Guimas Almeida, date: 12/05/2017, view: 135, Q&A: 0
    [Abstract] The established primary trigger of Alzheimer’s disease’s is β-amyloid (Aβ) (Mucke and Selkoe, 2012). Amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at endosomes also depends on its recycling back to the plasma membrane (Koo et al., 1996; Ubelmann et ...
    A Novel Protocol to Quantitatively Measure the Endocytic Trafficking of Amyloid Precursor Protein (APP) in Polarized Primary Neurons with Sub-cellular Resolution
    Authors:  Florent Ubelmann, Tatiana Burrinha and Cláudia Guimas Almeida, date: 12/05/2017, view: 86, Q&A: 0
    [Abstract] Alzheimer’s disease’s established primary trigger is β-amyloid (Aβ) (Mucke and Selkoe, 2012). The amyloid precursor protein (APP) endocytosis is required for Aβ generation at early endosomes (Rajendran and Annaert, 2012). APP retention at endosomes depends on its sorting for degradation in lysosomes (Haass et al., 1992; Morel et al. ...
    Notch Ligand Binding Assay Using Flow Cytometry
    Authors:  Shweta Varshney and Pamela Stanley, date: 12/05/2017, view: 156, Q&A: 0
    [Abstract] Notch signaling is an evolutionarily conserved signaling pathway that plays an indispensable role during development, and in the maintenance of homeostatic processes, in a wide variety of tissues (Kopan, 2012; Hori et al., 2013). The multifaceted roles of Notch signaling are stringently regulated at different levels. One of the most ...
    Formalin Murine Model of Pain
    [Abstract] Pain research is mostly based on experimental assays that use animal models, which may allow deciphering the physiopathology of this condition and to propel drug discovery. The formalin nociception test is considered one of the most predictive approaches to study acute pain in rodents. This test permits monitoring pain-related responses (i.e. ...
    GC/MS-based Analysis of Volatile Metabolic Profile Along in vitro Differentiation of Human Induced Pluripotent Stem Cells
    [Abstract] Human induced pluripotent stem cells (hiPSCs) are a promising tool in cell-based therapies for degenerative diseases. A safe application of hiPSCs in vivo, requires the detection of the presence of residual undifferentiated pluripotent cells that can potentially cause the insurgence of teratomas. Several studies point out that metabolic ...
    Mandibular Explant Assay for Investigating Extrinsic Stimuli on Bone and Cartilage Development
    Authors:  Jamie Jiang, Jessica W. Bertol and Walid D. Fakhouri, date: 12/05/2017, view: 139, Q&A: 0
    [Abstract] A major issue in developmental biology is to determine how embryonic tissues respond to molecular signals in a timely manner and given the position-restricted instructions during morphogenesis, of which Meckel’s cartilage is a classical example. The ex-vivo explant model is a practical and convenient system that allows investigation of ...
    [Bio101] RNA Extraction from Wistar Rat Cochlea for qRT-PCR
    Authors:  Janaína Cândida Rodrigues and Rubens Vuono de Brito Neto, date: 12/05/2017, view: 87, Q&A: 0
    [Abstract] Otology research has developed considerably in recent years and molecular analysis is crucial to identify metabolic pathways and therapeutic targets. However, the structure of the cochlea limits the amount of cell mass, and special care is required for RNA extraction. Studies applying this technique to the cochlea are scarce in the literature, and ...