Medium-throughput image-based phenotypic siRNA screen to unveil the molecular basis of B cell polarization

基于图像的中通量表型 siRNA 筛选揭示 B 细胞极化的分子基础

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作者:Dorian Obino, Mathieu Maurin, Florent Dingli, Damarys Loew, Aurianne Lescure, Emmanuel Terriac, Christel Goudot, Odile Malbec, Danielle Lankar, Maria-Isabel Yuseff, Ana-Maria Lennon-Duménil, Hélène D Moreau

Abstract

Cell polarity is an essential and highly conserved process governing cell function. Cell polarization is generally triggered by an external signal that induces the relocation of the centrosome, thus defining the polarity axis of the cell. Here, we took advantage of B cells as a model to study cell polarity and perform a medium-throughput siRNA-based imaging screen to identify new molecular regulators of polarization. We first identified candidates based on a quantitative proteomic analysis of proteins differentially associated with the centrosome of resting non-polarized and stimulated polarized B cells. We then targeted 233 candidates in a siRNA screen and identified hits regulating the polarization of the centrosome and/or lysosomes in B cells upon stimulation. Our dataset of proteomics, images, and polarity indexes provides a valuable source of information for a broad community of scientists interested in the molecular mechanisms regulating cell polarity.

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