Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages

评估小鼠成年小胶质细胞和骨髓来源巨噬细胞中的mRNA翻译

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作者:Ignazio Antignano ,Lily Keane ,Melania Capasso

Abstract

Protein synthesis, or mRNA translation, is the biological process through which genetic information stored in messenger RNAs is encoded into proteins. Here, we present an optimized protocol for assessing the translation rate in mouse adult microglia and cultured bone-marrow-derived macrophages. We describe steps for isolating cells, treating them with a puromycin-analog probe, and fluorescently labeling the puromycylated-polypeptide chains. We then detail their quantification by flow cytometry or with a fluorescent plate reader. For complete details on the use and execution of this protocol, please refer to Keane et al. (2021).1.

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