Journal: iScience
Article Title: Chronic stress antagonizes formation of stress granules
doi: 10.1016/j.isci.2025.114556
Figure Lengend Snippet: Chronic stress-pre-incubated cells fail to disassemble polysomes by acute stress U2OS cells were subjected to treatment with 100 μM SA for 1 h after pre-incubation with 10 μM SA, 1 μM Tg (thapsigargin), 60 μM CCCP (carbonyl cyanide m -chlorophenyl hydrazone), or HBSS for 24 h. Unstressed cells (NT) were used as a control. (A) Cells were pulsed with puromycin and emetine for 5 min and lysed. Cell lysates were subjected to western blotting using antibodies for puromycin, p -eIF2α, total eIF2α, G3BP1, G3BP2, caprin1, USP10, and β-actin. Three blots were taken for each of the three biological replicates (independent experiments). The same colors of the circle dot in the graph are from the same independent experiment. Results are mean ± S.E.M. ( n = 3). p values were assessed using a one-way ANOVA ( p ∗ <0.05, p ∗∗ <0.01, p ∗∗∗ <0.001, p ∗∗∗∗ <0.0001). (B–F) Polysome profiles from U2OS cells. NT, black; 1 h SA, red; (B) 24 h pre-incubation of SA, blue; 1 h SA with 24 h pre-incubation of SA, green. (C) 24 h pre-incubation of Tg, blue; 1 h SA with 24 h pre-incubation of Tg, green. (D) 24 h pre-incubation of CCCP, blue; 1 h SA with 24 h pre-incubation of CCCP; green. (E) 24 h pre-incubation of HBSS, blue; 1 h SA with 24 h pre-incubation of HBSS; green. (F) The polysome/monosome ratio was calculated with normalization by each of NT as a control.
Article Snippet: Rabbit Polyclonal USP10 antibody , Bethyl Laboratories Inc. , A300-900A; RRID: AB_625312.
Techniques: Incubation, Control, Western Blot
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