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The media was changed every 48-72 hours by discarding the media, washing the cells either once (for J774A.1 cell lines) or three times (for Caco-2 cell lines) in pre-heated 1 x phosphate buffered saline (PBS), and subsequently adding new, pre-heated DMEM with appropriate supplements

Four potential mechanisms

Steady state of NAC was expected to be attained after seven consecutive doses administered up to the morning of Day 6 with a of 12 h on the basis of mean half-life values of up to 14 h found in previous studies of the same dosage form