Mice were anesthetized​ with isoflurane and suspended using dental floss to allow oral access. The tongue was gently encouraged out of the mouth and held in place with a pipette tip to ensure the mouse was unable to swallow. Fifty microliters of control solution or treatment was delivered into the trachea via a standard lab pipette (20–200 μL). The mouse was held in position until its breathing returned to normal.

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