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. 2019 Feb 15;20(4):832. doi: 10.3390/ijms20040832

Figure 3.

Figure 3

Evaluation of death pathways triggered by fatty acids on Caco-2 cells. (A) The presence of apoptotic/necrotic cells was investigated after AnnexinV/Propidium Iodide (PI) staining and flow cytometry analysis. Representative plots of AnnexinV (X-axis/PI (Y-axis) staining of Caco-2 cells are shown. (B) Caspase 3/7 activation was evaluated by a luminometric assay. Cells were exposed for 8, 16 or 24 h to 150 µM palmitic acid, 250 µM palmitoleic acid and 300 µM sapienic acid (at these concentrations 75–80% reduction of cell viability is observed). (C) Caco-2 cells were pretreated for 3 h with the pan-caspase inhibitor Z-VAD or with the necroptosis inhibitor necrostatin-1 (Nec-1) and then treated with 150 µM palmitic acid, 250 µM palmitoleic acid and 300 µM sapienic acid. Cell viability was measured after 48 h. Results are means of three different experiments each performed in triplicate and they represent the percentage of control values obtained from cultures grown in the absence of fatty acid. SD never exceeded 15%. (*** p < 0.001; **** p < 0.0001).