| In vitro Differentiation of Murine Innate Lymphoid Cells from Common Lymphoid Progenitor Cells
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Author:
Date:
2016-03-20
[Abstract] Subtypes of innate lymphoid cells (ILC), defined based on their cytokine secretion profiles and transcription factor expression, are important for host protection from pathogens and maintaining tissue homeostasis. ILCs develop from common lymphoid progenitors (CLP) in the bone marrow. Using the methods described here, we have previously shown that loss of the transcriptional regulator TOX (Thymocyte-selection associated HMG-box protein) leads to specific changes in ILC development and differentiation. Here, we describe how to obtain ILCs from in vivo isolated CLP grown in vitro.
[摘要] 先天淋巴细胞（ILC）的亚型，基于其细胞因子分泌谱和转录因子表达来定义，对于宿主保护免于病原体和维持组织内稳态是重要的。 ILC从骨髓中的普通淋巴祖细胞（CLP）发展而来。 使用本文所述的方法，我们以前显示转录调节器TOX（胸腺细胞选择相关的HMG框蛋白）的损失导致ILC发展和分化的具体变化。 在这里，我们描述了如何从体内分离的CLP获得ILC。
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| Preparation of Recombinant Galectin-3 for Cancer Studies
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Author:
Date:
2016-01-05
[Abstract] Galectin-3 is a member of a class of proteins termed Galectins, characterized by their ability to bind glycans containing β-galactose (Cummings and Liu, 2009). Galectin-3 binds preferentially to proteoglycans terminating with N-acetyllactosamine (LacNAc) chains (i.e., tandem repeats of galactose) (Newlaczyl and Yu, 2011). Galectin-3 is unique among the galectins in its chimeric structure. It shares a conserved carbohydrate recognition domain (CRD) with the other galectins, but has a long amino-terminal tail that is thought to be involved in protein aggregation. It can also form homodimers through its CRD (Cummings and Liu, 2009). Galectin-3 has been found to have diverse functions in tumorigenesis including: signaling, apoptosis inhibition, immune suppression, cell growth, and ...
[摘要] Galectin-3是称为Galectins的一类蛋白质的成员，其特征在于它们结合含有β-半乳糖的聚糖的能力（Cummings和Liu，2009）。 Galectin-3优先结合以N-乙酰乳糖胺（LacNAc）链（即半乳糖的串联重复）终止的蛋白聚糖（Newlaczyl和Yu，2011）。 Galectin-3在其嵌合结构中的半乳凝素中是独一无二的。它与其他半乳凝素共享保守的碳水化合物识别结构域（CRD），但具有被认为参与蛋白质聚集的长氨基末端尾部。它也可以通过CRD形成同型二聚体（Cummings和Liu，2009）。已经发现Galectin-3在肿瘤发生中具有多种功能，包括：信号转导，凋亡抑制，免疫抑制，细胞生长和转移等。 ...
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| Helicase Assays
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Author:
Date:
2014-03-20
[Abstract] Helicases are a class of enzymes which are motor proteins using energy derived from ATP hydrolysis to move directionally along a nucliec acid phosphodiester backbone (such as DNA, RNA and DNA-RNA hybrids) and separate two annealed nucleic acid strands. Many cellular processes, such as transcription, DNA replication, recombination and DNA repair involve helicase activity. Here, we provide a protocol to analyze helicase activities in vitro. In this protocol, the DNA helicase protein Merkel cell polyomavirus large T-antigen was expressed in the mammalian cell line HEK293 and immoblized on an IgG resin. The helicase assay is performing while the protein is immoblized on IgG resin.
[摘要] 解旋酶是一类酶，其是使用来自ATP水解的能量沿着核酸磷酸二酯主链（例如DNA，RNA和DNA-RNA杂交体）定向移动并分离两条退火的核酸链的运动蛋白。 许多细胞过程，例如转录，DNA复制，重组和DNA修复涉及解旋酶活性。 在这里，我们提供了一个协议，以分析解旋酶活性在体外。 在该协议中，DNA解旋酶蛋白Merkel细胞多瘤病毒大T抗原在哺乳动物细胞系HEK293中表达并固定在IgG树脂上。 进行解旋酶测定，同时蛋白质固定在IgG树脂上。
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