Figure 2.

Analysis of cell viability and oxidative stress following exposure to SA4503, PRE084, and (+)-PTZ. (A) These 661W cells were exposed to tBHP (55 µM) to induce oxidative stress in the presence/absence of Sig1R ligands SA4503 (25, 50, and 100 µM), PRE084 (25, 50, and 100 µM), or (+)-PTZ (25, 50, and 100 µM) for 24 hours. Cell viability was measured using the MTT assay. Independent experiments were performed three times with four repetitions per assay. (B) These 661W cells were seeded on coverslips for 18 hours, after which they were exposed 2 hours to tBHP (55 µM) in the presence or absence of SA4503 (50 and 100 µM), PRE084 (50 and 100 µM), or (+)-PTZ (50 µM). They were incubated with CellROX Green Reagent to detect ROS; green fluorescent signals indicating ROS were visualized by epifluorescence. DAPI was used to label nuclei (blue). Calibration bars = 100 µm. (C) Fluorescence intensity was quantified by ImageJ. The experiment was performed three times with three repetitions of the assay. One-way ANOVA, significance is indicated in reference to tBHP group: *P < 0.05; **P < 0.01; ****P < 0.0001.