Figure 3: SFN prevents GM-induced apoptosis in LLC-PK1 cells. (a) Representative images (20x) obtained by fluorescence microscopy showing necrotic and apoptotic LLC-PK1 cells after 72 h of exposure to GM. Necrotic cells were stained with PI and apoptotic cells were stained with the annexin V-FITC probe. Cells were incubated with (SFN and SFN + GM groups) or without (CT and GM groups) 5 μM SFN for 24 h, followed by the addition of 8 mM GM (GM), 5 μM SFN (SFN), 5 μM GM (SFN + GM), or medium (CT) for 72 h. All of groups showed similar PI and annexin V-FITC fluorescence, but the cell population was different between them (bright field photographs). (b) Fluorescence intensity corrected for cell number (measured by trypan blue exclusion assay, TBS cells/well + viable cells/well in Table 1) of each group. After pictures taking, fluorescence intensity was measured as was explained in Section 2.3.Data are of 3 independent experiments. , versus CT; versus GM.