ArticleMethods in molecular biology (Clifton, N.J.)2027
Probing the Role of RNA Helicases in Modulating Stress Granules, RNA Localization, and P-Body Docking.
Article in Methods in molecular biology (Clifton, N.J.), 2027. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Stress granules (SGs) are cytoplasmic, membrane-less ribonucleoprotein (RNP) granules that form in response to cellular stress following the inhibition of translation initiation. Their assembly, composition, and crosstalk with processing bodies (P-bodies) are mediated by protein-protein, RNA-protein, and RNA-RNA interactions. DEAD/DEAH-box RNA helicases are key regulators of stress granules. Here, we present standardized protocols for (i) immunofluorescence of stress granule and P-body scaffold proteins and helicases, (ii) single-molecule RNA fluorescence in situ hybridization (smRNA FISH), and (iii) quantitative image analysis using Imaris software, including single-cell and nuclear segmentation, stress granule and P-body surface rendering, docking interaction analysis, and RNA spot detection.
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