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Total cells from the BAL were centrifuged and counted using Countess II Automated Cell Counter (Invitrogen, Carlsbad, CA) as previously described ( Morphometric assessment In order to determine the effects of ENDS exposure on the development of emphysema, we did a morphometric assessment according to the previous protocol ( Immunohistochemical, immunofluorescent, and histology analysis The mouse lungs were inflated with 1% low melting-point agarose to 25 cm of fixative pressure and then fixed with 4% neutral buffered formalin for 48 hours

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