Fig. 5

Effect of PJ-frozen storage on mitochondrial function in MDA cells. PJ was stored at -20 °C for 30 days, then MDA cells were incubated 24 h with DMEM medium containing different concentrations of frozen PJ. a Laser scanning confocal microscopy (LSCM) imaging of MDA treated with the reactive oxidant species probe DCF. Left panels show representative images of MDA cells treated with different PJ concentration. The histogram on the right shows statistical analysis of the DCF pixel intensity per cell normalized to control. b Representative Normalized Cell Index (NCI) kinetics of cells exposed to increase of PJ concentration. Cells were grown in complete DMEM medium for approximately 24 h after seeding, then medium was changed, and cells were treated for 5 days with DMEM medium containing different concentrations of PJ. Each cell index value was normalized just before this starting point. c The histogram displays NCI values at different days and different PJ-concentration of MDA cells. All data are presented as mean ± SEM of two independent experiments. ** p < 0.01; *** p < 0.001; **** p < 0.0001. Statistical significances are referred to the controls