TLR signaling in C57BL/6J TLR2,4,7-tolerant splenocytes over PBS-treated counterpart.. Mus musculus

Murine splenocytes were isolated from the spleens of C57BL/6J mice and were pretreated in vitro for three days in the presence of Pam3CSK4 (1 μg/ml), high pure LPS from E.coli O111:B4 (100 ng/ml) and R848 (5 μg/ml). PBS-treated splenocytes 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. Splenocytes from 6 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.