Fig. 4: Relationship between SG formation and the translation-controlling pathways.

A–D Protein expression profile of the ISR and mTOR pathways, in hOLs. Time course experiment of hOLs treated in vitro with SA, LG, or NG conditions. Cells were labeled with p-eIF2α (A) (n = 3 independent biological replicates) and p4E-BP1 (C) (n = 5 independent biological replicates) antibodies. The intensity of these markers was quantified per cell and normalized to the control condition. The left part of the X axis shows up to 10 h—the right part of the X axis from 1 to 4 days under the indicated treatments. Representative confocal images of hOLs are shown for p-eIF2α (B) and p4E-BP1 (D) at 4 h. E Time course experiment of hOLs exposed to SA conditions, in the presence or absence of Sephin1 or ISRIB (n = 5). Statistical analysis was performed between SA and SA+drug groups. F Quantitative analysis of hOLs exposed to NG conditions, in the presence or absence of Sephin1 or ISRIB for 4 h (n = 3). G, H Expression profile of p4E-BP1 protein in hOLs exposed to Ctrl or NG conditions in the presence or absence of Torin1 inhibitor for 4 h (n = 4). The intensity of these markers was quantified per cell and normalized to control condition. Representative confocal images of cells after 4 h are shown in (H). I Time course experiment showing the percentages of SG-positive hOLs, when exposed to NG conditions, in the presence or absence of Torin1 inhibitor (n = 4). Statistical analyses were performed between Ctrl and Ctrl+drug or NG and NG + drug groups. Scale bars, 10 µm. Merge pictures display DAPI (blue), O4 (pink), and the marker of interest (green). The percentages of SG-positive cells were assessed by G3BP1. Graphs display arbitrary units (arb. units). Each dot in the graphs corresponds to an independent biological replicate. All data are expressed as mean values ± SEM, analyzed by paired two-tailed Student’s t test (A, C, E, I) or by one-way ANOVA (F, G) followed by Bonferroni’s multiple comparisons correction. All significant P values are indicated; ns or unlabeled not significant. Ctrl optimal media, SA sodium arsenite, LG/NG low or no glucose. Source data are provided as a Source Data file.