Fig. 3: Relationship between SGs and ATP levels and protein synthesis under acute and chronic stress conditions.

A ATP levels in hOLs treated with Ctrl, SA, or NG conditions at different time points. The luminescence (ATP) of each condition was normalized per cell (n = 3). No significant differences were found between NG and NG + TNFα + IFNγ conditions. B, C Quantitative analysis of protein synthesis of hOLs treated with Ctrl, SA, NG, and CHX at the indicated time. Protein synthesis was assessed by immunofluorescent signal intensity following labeling with L-homopropargylglycine (HPG) and Click-iT assay kit. Representative confocal images of protein synthesis in hOLs using HPG staining are shown in (C). Cells were cultured under Ctrl or stress conditions for 4 h. For each condition, the specific number of biological samples tested is shown in the graph. Control cells show high signal intensity (normal translation) whereas cells under stress conditions show a significant decrease in signal intensity (translation inhibition). Scale bars, 10 µm. Merge pictures display DAPI (blue), O4 (pink), and HPG (green). (D) Quantification of the percentage of cells with SGs during continual exposure to Ctrl, NG, SA, and NG + TNFα + IFNγ conditions in combination with puromycin or CHX for 4 and 24 h. For each condition, the specific number of biological samples tested is shown in the graph. Each dot in the graphs corresponds to an independent biological replicate. All data are expressed as mean values ± SEM, analyzed by one-way ANOVA followed by Bonferroni’s multiple comparisons correction. All significant P values are indicated; ns or unlabeled not significant. Ctrl optimal media, SA sodium arsenite, NG no glucose, CHX cycloheximide. Source data are provided as a Source Data file.