Characterization of RACK1-depleted mammalian cells by a palette of microscopy approaches reveals defects in cell cycle progression and polarity establishment

Exp Cell Res. 2023 Jun 30:113695. doi: 10.1016/j.yexcr.2023.113695. Online ahead of print.ABSTRACTThe Receptor for Activated C Kinase 1 (RACK1) is an evolutionarily conserved scaffold protein involved in the regulation of numerous cellular processes. Here, we used CRISPR/Cas9 and siRNA to reduce the expression of RACK1 in Madin-Darby Canine Kidney (MDCK) epithelial cells and Rat2 fibroblasts, respectively. RACK1-depleted cells were examined using coherence-controlled holographic microscopy, immunofluorescence, and electron microscopy. RACK1 depletion resulted in decreased cell proliferation, increased cell area and perimeter, and in the appearance of large binucleated cells suggesting a defect in the cell cycle progression. Our results show that the depletion of RACK1 has a pleiotropic effect on both epithelial and mesenchymal cell lines and support its essential role in mammalian cells.PMID:37393981 | DOI:10.1016/j.yexcr.2023.113695

Source: Experimental Cell Research - July 2, 2023 Category: Cytology Authors: Vera Chvalova Vignesh Venkadasubramanian Zuzana Klimova Jana Vojtova Oldrich Benada Ondrej Vanatko Tomas Vomastek Tomas Grousl Source Type: research