Fig. 4

Time-of-day immune cell activation. PBMLs were isolated at 6 am or 6 pm and stimulated for 2 h with LPS/anti-CD3/28. Then mRNA was purified and subject to RNA-SEQ analysis. a Venn diagrams were generated using all the significantly expressed genes in each comparison. b Genes that were significantly regulated by LPS at either time point in RA were subjected to Edge set analysis (pathway-result network diagram shown). The Edges that contribute to the Edge set are labelled in red. c AM and d PM LPS-regulated genes were compared (healthy vs RA) (shown as Venn diagrams). Genes that were unique to RA at each time point (from c and d) were analysed using the Reactome database (AM genes (e) and PM genes (f)). g LPS-stimulated phospho-proteome analysis. Significant sites are shown in red (unregulated by LPS and blue (downregulated by LPS). Motif analysis of LPS-regulated sites shown below. h Luminex assay for phospho-p38, HSP27, STAT3, ATF2 and fluorescence values was log transformed (mean and SD shown) and analysed using a repeated measures ANOVA