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Fig. 5. Changes of intracellular ROS and free Ca²⁺ levels in Hend cells checked by confocal analysis. Cells were exposed for 15, 30, 60, 180 minutes and 24 hours to 2.0 µM HypF-N pre-fibrillar aggregates and then fixed with 2.0% paraformaldehyde. ROS were obtained by incubating for 10 minutes the exposed cells in the presence of the redox fluorescent probe and by measuring the fluorescence of CM-H2 DCFA. Intracellular free Ca²⁺ levels were revealed by incubating the exposed cells for 15 minutes in the presence of the fluorescent dye Fluo-3AM. The data are reported as percentages of the values determined at time 0 and are expressed as means ±s.d. of four experiments each carried out in duplicate. For technical details see under Materials and Methods.

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