Lovejoy Madeline, Mucci Claudio A, Rabino Agustín, Garcia-Mata Rafael
Department of Molecular, Cellular, and Developmental Biology, University of Toledo, Toledo, OH 43606, USA.
Department of Molecular, Cellular, and Developmental Biology, University of Toledo, Toledo, OH 43606, USA.
STAR Protoc. 2025 Sep 19;6(3):104079. doi: 10.1016/j.xpro.2025.104079. Epub 2025 Sep 10.
Polarized epithelial cells form a tightly packed monolayer where individual cells are connected by cell-cell junctions, including adherens junctions (AJs) and tight junctions (TJs). Here, we present a protocol for quantifying the horizontal and vertical distribution of junctional proteins in confluent, fixed epithelial cells between different experimental conditions. We describe steps for sample preparation, immunofluorescence, and image acquisition. We also provide detailed instructions on how to use Fiji/ImageJ and RStudio for this procedure without any prior experience necessary. For complete details on the use and execution of this protocol, please refer to Rabino et al..
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，包括黏附连接（AJs）和紧密连接（TJs）。在此，我们提供了一种方案，用于量化在不同实验条件下汇合的固定上皮细胞中连接蛋白的水平和垂直分布。我们描述了样品制备、免疫荧光和图像采集的步骤。我们还提供了关于如何在无需任何先验经验的情况下使用Fiji/ImageJ和RStudio进行此过程的详细说明。有关此方案的使用和执行的完整详细信息，请参考拉比诺等人的研究。
