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At the end of the experiment, the cells were washed once with Hanks balanced salt solution (HBSS) and cell viability was assessed by adding 20 L of the CellTiter 96 AQueous One Solution reagent (Promega, Fitchburg, WI) to 100 l HBSS, followed by incubation for 60 min at 37 C, and the absorbance was measured at 490 nm using the Infinite 200 PRO plate-reader (Tekan, Switzerland)