Fig. 2

CLS determination using the PI assay (PICLS with human cells). a CLS determination by PI assay with 2,5-AM treatment. Cells were seeded in a 96-well plate with PI (5 µg/ml) dissolved in the D10 medium. Fluorescence reading was taken on the fifth day after seeding. b CLS determination by the PI assay with rapamycin treatment, a similar experimental procedure was performed as in (a). c Schematic diagram illustrating the experimental procedure of the cell viability assay. DMSO (d) and water-soluble (e) compounds screening using PICLS assay determines the potential anti-aging compounds that extend cellular lifespan. 50 nM rapamycin (d) and 20 mM 2,5-AM (e) were used as positive controls. Graphs depict data from Day 6 results. f CLS determination by cell viability assay with compounds found positive in (d–e). a–b, f were analyzed by ordinary one-way ANOVA followed by Dunnett’s multiple comparisons tests. Results are plotted as mean ± SD. *P ≤ 0.05; **P ≤ 0.01