Lovejoy Madeline, Mucci Andoni, Rabino Agustín, Garcia-Mata Rafael
Department of Biological Sciences, University of Toledo, Toledo, OH 43606, USA.
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bioRxiv. 2025 May 5:2025.05.01.651687. doi: 10.1101/2025.05.01.651687.
Polarized epithelial cells form a tightly packed monolayer where individual cells are connected by cell-cell junctions, including tight junctions (TJ) and adherens junctions (AJ). Here, we present techniques for quantifying the horizontal and vertical distribution of junctional proteins in confluent, fixed epithelial cells. This approach is utilized to evaluate variations in the intensity and localization of the proteins that compose the AJ and TJ under different experimental conditions. Although our protocol is optimized for Madin-Darby Canine Kidney (MDCK) cells, it is adaptable to any cell line capable of forming cell-cell junctions. For complete details on the use and execution of this protocol, please refer to Rabino et al. (2024). .
极化上皮细胞形成紧密堆积的单层，其中单个细胞通过细胞间连接相连，包括紧密连接（TJ）和黏附连接（AJ）。在这里，我们展示了用于量化汇合的固定上皮细胞中连接蛋白水平和垂直分布的技术。该方法用于评估在不同实验条件下组成AJ和TJ的蛋白质的强度和定位变化。尽管我们的方案是针对犬肾上皮细胞（MDCK）优化的，但它适用于任何能够形成细胞间连接的细胞系。有关此方案的使用和执行的完整详细信息，请参考拉比诺等人（2024年）。
