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    Protocols in Current Issue
    High Dimensional Functionomic Analysis of Human Hematopoietic Stem and Progenitor Cells at a Single Cell Level
    Authors:  Thomas Luh, Kimberly Lucero, Wenji Ma, Jaeyop Lee, Yu Jerry Zhou, Yufeng Shen and Kang Liu, date: 05/20/2018, view: 132, Q&A: 0
    The ability to conduct investigation of cellular transcription, signaling, and function at the single-cell level has opened opportunities to examine heterogeneous populations at unprecedented resolutions. Although methods have been developed to ...
    High Dimensional Functionomic Analysis of Human Hematopoietic Stem and Progenitor Cells at a Single Cell Level
    Authors:  Thomas Luh, Kimberly Lucero, Wenji Ma, Jaeyop Lee, Yu Jerry Zhou, Yufeng Shen and Kang Liu, date: 05/20/2018, view: 132, Q&A: 0
    [Abstract] The ability to conduct investigation of cellular transcription, signaling, and function at the single-cell level has opened opportunities to examine heterogeneous populations at unprecedented resolutions. Although methods have been developed to evaluate high-dimensional transcriptomic and proteomic data (relating to cellular mRNA and protein), ...
    Purification of Total RNA from DSS-treated Murine Tissue via Lithium Chloride Precipitation
    Authors:  Emilie Viennois, Anika Tahsin and Didier Merlin, date: 05/05/2018, view: 190, Q&A: 0
    [Abstract] We have developed a protocol to purify RNA from DSS (Dextran Sulfate Sodium)-treated mouse tissues. This method, which prevents downstream inhibition of q-RT-PCR observed in DSS-treated tissues, relies on successive precipitations with lithium chloride.
    Nab Escaping AAV Mutants Isolated from Mouse Muscles
    Authors:  Zheng Chai, R. Jude Samulski and Chengwen Li, date: 05/05/2018, view: 161, Q&A: 0
    [Abstract] Neutralizing antibodies (Nabs) are a major challenge in clinical trials of adeno-associated virus (AAV) vector gene therapy, because Nabs are able to inhibit AAV transduction in patients. We have successfully isolated several novel Nab-escaped AAV chimeric capsids in mice by administrating a mixture of AAV shuffled library and patient serum. These ...
    3D Co-culture System of Tumor-associated Macrophages and Ovarian Cancer Cells
    Authors:  Lingli Long, Mingzhu Yin and Wang Min, date: 04/20/2018, view: 476, Q&A: 0
    [Abstract] Ovarian cancer is fairly unique in that ovarian carcinoma cells can detach and spread directly through peritoneal cavity. It has been unclear, however, how detached cancer cells survive in the peritoneum and form spheroid structure. We have recently reported that there is a strong correlation between Tumor-associated macrophages (TAMs)-associated ...
    Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
    Authors:  Jorge Luis Ayala-Lujan and Fernando Ruiz-Perez, date: 04/20/2018, view: 309, Q&A: 0
    [Abstract] Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory ...
    Imaging Cytokine Concentration Fields Using PlaneView Imaging Devices
    Authors:  Alon Oyler-Yaniv and Oleg Krichevsky, date: 04/05/2018, view: 310, Q&A: 0
    [Abstract] We describe here a method to visualize concentration fields of cytokines around cytokine-secreting cells. The main challenge is that physiological cytokine concentrations can be very low, in the pico-molar range. Since it is currently impossible to measure such concentrations directly, we rely on cell’s response to the cytokines–the ...
    Intravenous Labeling and Analysis of the Content of Thymic Perivascular Spaces
    Authors:  Roland Ruscher and Kristin A. Hogquist, date: 03/05/2018, view: 419, Q&A: 0
    [Abstract] Following development in the thymus, T cells are thought to exit into the periphery predominantly through perivascular spaces (PVS). This exit route is used by conventional T cells, and likely also applies to unconventional T cell subsets, such as precursors of CD8αα and TCRγδ intraepithelial lymphocytes, regulatory T cells and natural killer T ...
    Mono Sodium Urate Crystal-induced Peritonitis for in vivo Assessment of Inflammasome Activation
    Authors:  Marianne R. Spalinger and Michael Scharl, date: 03/05/2018, view: 558, Q&A: 0
    [Abstract] Due to its particulate material, mono-sodium urate (MSU) crystals are potent activators of the NOD-like receptor NLRP3. Upon activation, NLRP3 induces the formation of inflammasome complexes, which lead to the production and release of mature IL-1β. Bioactive IL-1β is a potent activator of innate immune responses and promotes recruitment of ...
    Quantification of Bacterial Attachment to Tissue Sections
    Authors:  Batya Isaacson, Tehila Hadad, Gilad Bachrach and Ofer Mandelboim, date: 03/05/2018, view: 495, Q&A: 0
    [Abstract] Here we describe a method to test bacterial adhesion to paraffin embedded tissue sections. This method allows examining binding of different bacterial strains, transfected with a fluorescent protein reporter plasmid to various tissues, to better understand different mechanisms such as colonization. This assay provides a more physiological context ...
    Murine Pancreatic Islets Transplantation under the Kidney Capsule
    [Abstract] Type 1 diabetes (T1D) is an autoimmune disease caused by the lack of insulin-producing pancreatic beta cells leading to systemic hyperglycemia. Pancreatic islet transplantation is a valid therapeutic approach to restore insulin loss and to promote adequate glycemic control. Pancreatic islet transplantation in mice is an optimal preclinical model ...