Fig. 4

RT-qPCR validation of differentially expressed candidate loci following various LPS dose challenges. This figure illustrates the RT-qPCR expression of a tolerisable genes (TNF-α) and HERV/MaLR elements (121601901-HERV0116uL and 081146702-MALR1129uL) exhibiting a TNF-α–like pattern of expression and b non-tolerisable genes (IL10) and HERV/MaLR elements (070278702-MALR1045uL and 043166701-MALR1020uL) exhibiting an IL10–like pattern of expression. Expression was measured using mRNA derived from the stimulated and unstimulated PBMCs of the healthy volunteers used in the discovery microarray experiment (first column, (Aa and Ba)), mRNA derived from the stimulated and unstimulated PBMCs of 6 additional healthy volunteers managed in the same way (second column, (Ab and Bb)), and finally mRNA derived from the stimulated and unstimulated PBMCs of 5 additional healthy volunteers (third column, (Ac and Bc)). This last column includes additional IFN-γ dependant reversibility of tolerance, consisting of a 2 ng/mL LPS priming step overnight, followed by a 100 ng/mL IFN-γ stimulation step overnight, and finally the 100 ng/mL LPS stimulation step for 6 hours. All PCR reactions were performed in duplicate for each condition. Expression of the housekeeping genes PPIB and RPLP0 was monitored for normalisation. The fold change (FC) was determined using the 2^-ΔΔCt method. The final value of the unstimulated condition was arbitrarily set to one and other values scaled-up in order to provide a final relative differential expression (data were represented by a median and using the log2 scale). Statistically significant differences between two conditions are marked (wilkoxon signed rank test. **: p-value < 0.05 and * p-value < 0.1)