Figure 1. Microarray analysis of ciliating mouse tracheal epithelial cells. (.

A) Immunofluorescence staining showing representative images of GFP expression and ciliogenesis in FOXJ1:GFP MTEC cultures before establishment of the air-liquid interface (pre-ALI), at 4 days after ALI (ALI+4) and at 12 days after ALI (ALI+12). MTEC cultures were stained with antibodies against GFP, against glutamylated tubulin to mark cilia and basal bodies, and with DAPI to mark nuclei. GFP is expressed from the FOXJ1 promoter early during ciliogenesis, but is not expressed pre-ALI. At ALI+4, most FOXJ1:GFP+ cells are undergoing centriole formation but have not formed cilia. At ALI+12, most FOXJ1:GFP+ cells are forming cilia or have completed ciliogenesis. Scale bars, 5 µm. (B) FACS analysis of cells dissociated from a trachea from a wild type mouse (left panel) and a trachea from a FOXJ1:GFP-expressing mouse (right panel). The red and blue rectangles are representative gates used to sort GFP+ and GFP- populations, respectively. (C) Sorted cell populations were stained with DAPI (blue) and combined acetylated alpha-tubulin and gamma-tubulin antibodies (green) to detect cilia and basal bodies. After sorting, 90–95% of cells in the GFP+ population at ALI+12 had observable cilia and or amplified basal bodies, whereas <1% of cells in the GFP- population stained positive for these markers, although some had a single primary cilium (arrowhead).