Wild type (+/+) and RCAN1-deficient (-/-) BMMs were treated with 200 ng/ml P. aeruginosa LPS for various time points or left untreated (NT). The total RNA isolated from these cells was reverse transcribed to cDNA and subjected to real-time quantitative PCR for IRF3 (A), and IRF7 (B) gene expression. The gene expression was normalized to housekeeping control gene HPRT. Cell lysates were immunoblotted to measure IRF3, IRF7 and actin protein levels. Immunoblots are representative of three independent experiments (C). Densitometry analysis of IRF3 and IRF7 levels was normalized to actin (D, E), and data are presented as fold change (n = 3 ± SEM, **p<0.01, ****p<0.0001).