Bronchoalveolar lavage analysis.

Ten-day-old mice were injected with LPS, with or without the HDAC6 or pan-HDAC inhibitors as above, and bronchoalveolar lavage analysis (BAL) was performed at 24 h. Mouse lungs were lavaged 10 times with 200 µL of PBS after endotracheal cannulation. A 10 µL aliquot was loaded on a hemocytometer for cell number counting; 50 µL were used for quantification of total protein by a BCA assay, and a 200 µL aliquot was collected on a cytospin at room temperature; and Shandon Kwik Diff staining (commercially available from Thermo-Fisher) was performed following the manufacturer’s protocol on 300 cells/prep.