Mouse CpG stimulation and monocyte gene expression analysis

Female 8–10-week-old NOD and control B6.g7 mice (Jackson Laboratories, Ellsworth, ME, USA), 3–4 mice/group, were injected intravenously with 170 μl PBS + 30 μl 1,2-Dioleoyloxy-3-trimethylammonium propane (DOTAP; Roche, Indianapolis, IN, USA), with and without 5 μg CpG-A (2216; Invivogen, San Diego, CA, USA). After 6 h, spleen cells were harvested and Ly6C⁺ monocytes were sorted. Extracted RNA was sequenced and analysed, as previously described [16, 34, 35]. Details and other analysis of the unstimulated mouse samples used in this study were previously published [16]. The experiments were not replicated, with each mouse acting as a biological replicate.