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. 2023 Jan 31;51(3):1326–1352. doi: 10.1093/nar/gkad033

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Various RNA helicases re-localize to and co-precipitate in As[III]-induced SG. (A) Indirect immunofluorescence images (false-coloured and merged) of HeLa cells analysed for the subcellular localization of RNA helicases (magenta), which were repeatedly identified as SG-associated and that co-migrated with 5′ tsRNAs-containing RNPs (this work) and the ubiquitous SG marker, G3BP3 (cyan), during steady-state conditions. (B) Indirect immunofluorescence images of HeLa cells (false-coloured) analysed for the colocalization of various RNA helicases (magenta) with G3BP1 (cyan) after acute exposure to As[III] (0.3 mM for 1 h). (C) Western blotting of different candidate helicases on samples obtained during the steps of SG core enrichment according to (36) and after immunoprecipitation of eIF4A1 from enriched SG cores from HeLa cells were either acutely exposed to As[III] (0.5 mM for 1 h, acute, – recovery) or were incubated in fresh medium (+ recovery) after removal of As[III]. (D) Western blotting of different candidate helicases on samples obtained during the steps of SG core enrichment according to (36) and after immunoprecipitation of DDX3X from enriched SG cores extracted from Hela cells acutely exposed to As[III] (0.5 mM for 1 h, acute).