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In addition, cell viability assay (Trypan blue staining), apoptosis assay (annexin V), histological analysis (Hematoxylin and Eosin staining), confocal microscopy [for immunofluorescence detection of zona occludin 1 (ZO1) alternatively known as tight junction protein 1 (TJP1), SP-C, LysoTracker Green DND-25, epithelial sodium channel (ENaC)] and transmission electron microscopy (as described previously 28 ) was performed to characterize the alv-ALI model

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