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For immunostainings cells were blocked for 30 min in blocking buffer (10% GS, 0.3% Triton-X-100, 20 mM HEPES pH 7.4), then incubated with anti-p62 antibody (1:200, BD Bioscience, 610832), washed three times for 10 min in PBS, incubated with AF568-labeled goat-anti-mouse antibody, washed again three times in PBS for 10 min, and finally mounted using Shandon TM Immu-Mount TM containing Hoechst33342

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It was speculated that the loss of VIP-innervation to the airway diminished the amount of bronchodilation mediated by the nervous system

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