Fig. 5.

Effects of enhanced S1P turnover and FFA on overall oxidative stress and cytosolic hydrogen peroxide production in insulin-secreting INS1E cells. INS1E cells were incubated in the absence or presence of PA or OA (500 μM of each) for 24 h. (A) overall oxidative stress was measured by DCFDA method. (B) cytosolic hydrogen peroxide generation was estimated by expressing the hydrogen peroxide-sensitive fluorescence sensor HyPer-Cyto protein in the cytosolic compartment of cells and evaluation of fluorescence shift by the CellSens software at the Olympus fluorescence microscope (representative pictures from n = 3 individual experiments), Bars: 10 μm. Shown (A) are Means ± SEM from n = 3–6 independent experiments, each condition was measured in triplicates ANOVA followed by Bonferroni, ∗∗P < 0.01, ∗∗∗P < 0.001 versus untreated, #P < 0.05, ##P < 0.01, versus INS1E-ctr cells treated in the same way.