Figure 3.

Deletion of GR in airway epithelial cells of Ccsp-GR^−/− mice is associated with reduced responsiveness to Dex-induced circadian synchronization. A) Immunofluorescence images of lung sections from GR^WT (top) and Ccsp-GR^−/− mice (bottom) stained for GR (red) and cell nuclei (DAPI). Green indicates background/auto fluorescence. B) Images of lung sections from GR^WT (top) and Ccsp-GR^−/− mice (bottom) probed for GR mRNA via in situ hybridization with relative quantification of GR (Nr3c1) mRNA abundance from multiple lung sections (n = 4/genotype, analyzed with 2-tailed Student’s t test). C) PER2-LUC bioluminescence (photon counts/min) over multiple days from GR^WT (left) and Ccsp-GR^−/− (right) lung sections. Dex was administered at the indicated time (arrow). Plot is representative of 3 independent replicates. D) Relative quantification of Bmal1, Per2, Nr1d1, and Nr1d2 gene expression in whole lung (n = 4–7, 2-way ANOVA with Sidak’s multiple comparisons test within genotypes). For Nr1d1, 1 outlier was removed from the GR^WT group at CT12 after using the Grubbs’ test to detect outliers. Data represent means ± se. *P < 0.05, **P < 0.01, ***P < 0.001.