{{'Search' | translate}}
 

Purified Mouse Anti-β-Catenin Clone 14/Beta-Catenin (RUO)

纯化的小鼠抗β-联蛋白克隆14 /β-联蛋白

Company: BD
Catalog#: 610153
Bio-protocol()
Company-protocol()
Other protocol()

Lentiviral shRNA Screen to Identify Epithelial Integrity Regulating Genes in MCF10A 3D Culture
Author:
Date:
2016-12-05
[Abstract]  MCF10A 3D culture system provides a reductionist model of glandular mammary epithelium which is widely used to study development of glandular architecture, the role of cell polarity and epithelial integrity in control of epithelial cell functions, and mechanisms of breast cancer. Here we describe how to use shRNA screening approach to identify critical cell pathways that couple epithelial structure to individual cell based responses such as cell cycle exit and apoptosis. These studies will help to interrogate genetic changes critical for early breast tumorigenesis. The protocol describes a library of lentiviral shRNA constructs designed to target epithelial integrity and a highly efficient method for lentiviral transduction of suspension MCF10A cultures. Furthermore, protocols are ... [摘要]  MCF10A 3D文化系统提供了腺体乳腺上皮的还原剂模型,其广泛用于研究腺体结构的发育,细胞极性和上皮完整性在上皮细胞功能的控制中的作用以及乳腺癌的机制。在这里我们描述如何使用shRNA筛选方法来识别关键细胞通路,夫妇上皮结构到个别细胞的反应,如细胞周期退出和凋亡。这些研究将有助于询问对早期乳腺肿瘤发生至关重要的遗传变化。该协议描述了设计用于靶向上皮完整性的慢病毒shRNA构建体的文库和用于悬浮MCF10A培养物的慢病毒转导的高效方法。此外,提供的协议设置MCF10A 3D文化在Matrigel的形态和细胞反应研究通过结构化照明和共聚焦显微镜分析免疫染色的三维结构。
关键字: 3D文化,MCF10A,shRNA,上皮完整性,免疫荧光染色,3D成像,形态测量分析

[背景] 上皮细胞形成高度组织的组织结构,其提供物理支持和用于协调细胞信号传导的结构化支架。跨上皮结构的这种协调的信号传导对于上皮生物学是基本的;使得上皮细胞在调节器官大小,形状,功能和基于个体细胞的应答中的动态联合作用(Roignot等人,2013; Shamir和Ewald,2014)。上皮信号传导的联合指挥还提供了一种强有力的肿瘤抑制机制,通过将外部和内部有丝分裂信号门控到静止的上皮组织(Partanen等人,2013; Rejon等人 ...

Dissection and Staining of Mouse Brain Ventricular Wall for the Analysis of Ependymal Cell Cilia Organization
Author:
Date:
2016-03-20
[Abstract]  In the developing and mature central nervous system (CNS) the ventricular lumen is lined by the neuroepithelium and ependymal, respectively. These ventricular epithelia perform important functions related to the development, morphogenesis and physiology of the brain. In the mature CNS, ependyma constitutes a barrier between brain parenchyma and cerebro- spinal fluid (CSF). The most prominent feature of the apical surface of ependymal cells is the presence of multiple motile cilia that extend towards the ventricular lumen. The beating of cilia ensures the circulation of the CSF and its impairment leads to hydrocephalus. For an effective CSF flow, ciliary beating must be coordinated at the level of individual cells and at the tissue level. This coordination is achieved through the precise ... [摘要]  在发展中和成熟的中枢神经系统(CNS)中,心室腔分别由神经上皮和室管膜排列。这些心室上皮执行与脑的发育,形态发生和生理相关的重要功能。在成熟CNS中,室管膜构成脑实质和脑脊液(CSF)之间的屏障。室管膜细胞的顶面的最突出的特征是存在向心室腔延伸的多个运动性纤毛。纤毛的跳动确保CSF的循环,并且其损伤导致脑积水。对于有效的CSF流动,睫状细胞跳动必须在单个细胞水平和组织水平协调。这种协调是通过精确组织在室内平面内的纤毛定位来实现的。已经描述了关于室管膜细胞中纤毛的平面组织的两个主要特征(Mirzadeh等人,2010),并且它们都具有细胞和组织方面(Boutin等人 ,2014)。第一个,旋转极性,指睫毛跳动的方向。在细胞水平,所有纤毛在相同方向跳动(图1B,黑色箭头)。在组织水平,每个室管膜细胞协调其拍打的方向与相邻细胞的方向(图1C,灰色箭头)。第二个特征,翻译极性,对室管膜细胞是唯一的,是指簇中的纤毛聚集。在细胞水平,该簇相对于室管膜细胞的中心位移(图1B,红色箭头)。在组织水平,睫状簇的定位在相邻细胞之间协调(图1C)。在任一水平上改变任何这些极性都会损害CSF流动循环(Mirzadeh等人,2010; Boutin等人,2014; Guirao等人,/em,2010; Hirota等人,2010; ...

Comments