Murine CD3+ T-cells were immunomagnetically purified from the spleens of C57BL/6J mice and were pretreated in vitro for three days in the presence of R848 (5 μg/ml). Unmanipulated T-cells served as negative control. We used Qiagen Toll-like Receptor RT2 Profiler PCR Array kit to quantitate gene expression profiling of the TLR signaling pathway. Overall design: qPCR gene expression profiling. CD3+ T-cells from 5 mice were used and treated separately as indicated in the summary. Equal amount of total cDNA from each mouse (reverse-transcribed from 0.5 μg RNA) was used prior to gene expression analysis.