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Alternatively, cell viability was determined by Cell Titer-Glo (CTG) assay (Promega) plating 5000 cells/96-well in 100 l media, treatment of cells the next for 48 h followed by Cell Titer-Glo (CTG) assay (Promega) according to the manufacturers instructions

On the basis of the evidence presented above, the major objectives of the present study were (1) to determine the effects of GSHee on ventilatory performance in freely-moving adult rats, and (2) to determine whether the prior administration of GSHee is able to ameliorate the negative effects of fentanyl on breathing and gas-exchange in freely-moving adult rats