Exposure to stress and SGs formation.

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Exposure to stress and SGs formation. Exposure to stress and SGs formation. (A) SGs form in a concentration-depended manner in U2OS-G3BP1 and HEK-TIA1 cells following exposure to different concentrations of AS for 30 min (upper fluorescent images). TIA1 was visualized with FLAG antibodies (green), G3BP1 was visualized through its fluorescent GFP tag (green), and nuclei were counterstained with DAPI (blue). Control denotes untreated cells. Scale bar, 10 μm. Polysome profiles of HEK-TIA1 cells exposed to different AS concentrations (lower panel). 80S designates an assembled ribosome and 40S and 60S denote the ribosomal subunits. (B) mRNAs are sequestered in SGs. In situ hybridization (FISH) with combined probe for ADAMTS1, ADAMTS3, ADAMTS8, and ADAMTS15 transcripts (all also detected as m6A modified in the m6A-Seq) in U2OS-G3BP1 exposed for 30 min to AS. Scale bar, 10 μm. (C) Comparison between total mRNA from control HEK-TIA1 cells or exposed to 500 μM AS determined from RNA-Seq. Genes with significantly increased expression under stress are designated. R2 = 0.978, Pearson correlation coefficient. Source data are available for this figure. Maximilian Anders et al. LSA 2018;1:e201800113 © 2018 Ignatova et al.