Figure 4.

Human colon carcinoma cell line (Caco-2)/TC7 redox status. The accumulation of intracellular reactive oxygen species (ROS) was detected using the peroxide-sensitive fluorescent probe 2′,7′-dichlorofluorescein diacetate (DCFH-DA). Cells differentiated on transwell filters were untreated (C), treated with AAPH treated with Lactobacillus casei Shirota (LS) or pretreated with LS for 1.5 h and then treated with AAPH (LS + AAPH) for 2.5 h. After treatments, cells were washed three times with PBS and incubated with 10 µM DCFH-DA in HBSS. Redox status was measured by flow cytometry analysis (A) acquiring at least 10,000 events. The figure is representative of three independent assays. Fluorescence microscope analysis was performed on Caco-2/TC7 cells treated as previously described. The percentage of ROS positive cells with respect to total cells/field was calculated (B). At least 10 fields for each treatment were observed in three independent immunofluorescence assays. Data are reported as mean ± SD of percentage of DCFH-DA positive cells. Differences with P values <0.05 were considered significant and indicated within each parameter by **P < 0.01 vs C.