FIGURE LEGENDS:
Figure 1. miRNA expression signatures in PBS (control) and house
dust mite (HDM) exposed mouse lung. (A) Heatmap shows
unsupervised hierarchical clustering of nine differentially expressed
(DE) miRNAs in HDM vs. PBS exposed mouse lung at ZT0 analyzed by
Morpheus tool. (B) Pairwise comparison analysis performed by
nSolver 4.0 software showing DE miRNAs in HDM vs. PBS at ZT0.(C) Heatmap shows unsupervised hierarchical clustering of
thirty-eight DE miRNAs in HDM vs. PBS exposed mouse lung at ZT12
analyzed by Morpheus tool. (D) Pairwise comparison analysis
performed by nSolver 4.0 software showing DE miRNAs in HDM vs. PBS at
ZT12. Data are shown as fold change/ratio, P -value (P< 0.05) and FDR adjusted P -value < 1
(n=6/group/time point).
Figure 2. Representative data from differentially expressed
miRNAs and selected mRNA targets were analyzed by NanoString nCounter
analysis. (A) DE 9 miRNAs were significantly downregulated at
ZT0 and ZT12 pairwise comparison. We observed a time-of-day effect in
the expression of miRNAs in HDM vs. PBS exposed mouse lung. (B)Selected miRNA-mRNA targets predicted by Ingenuity Pathway Analysis were
further validated using NanoString mouse Myeloid Innate Immunity Panel
and qPCR analysis (il13 mRNA). Normalized counts data are
presented as analyzed by nSolver 4.0 software. (C)Bioinformatic analysis of DE miRNAs downregulated or upregulated in HDM
exposed group at ZT12 using DIANA-miRPath v3.0. KEGG pathways marked in
red color were pathways identified by miRPath analysis of DE miRNAs in
HDM vs. PBS at ZT12. Data are shown as mean ± SEM (n=6/group; * P< 0.05, ** P < 0.01, *** P <
0.001, significant compare to respective PBS control;# P < 0.05, # #P < 0.01, # # # P< 0.001, significant compared to PBS or HDM from ZT0; 2way
ANOVA with Tukey’s multiple comparison test).