Fig. 5: Innate immune cells derived from Psen2^N141I/+ mice are rhythmic despite suppressed expression of clock genes.

a, b Comparison of relative mRNA expression levels of clock genes between WT and KI/+ primary microglia (a; n = 4 for Cry1; n = 5 for other genes) (WT vs KI/+: ^###p = 0.0003, ^##p = 0.0011, ^##p = 0.0021, ^#p = 0.042, ^###p = 0.0001, and ^###p < 0.0001) and BMDM cultures (b) (n = 4 for Dbp, Rora and Csnk1a1; n = 5 for other genes) (WT vs KI/+: ^##p = 0.0022, ^##p = 0.008, ^#p = 0.0221, ^##p = 0.0049, ^#p = 0.0181, and ^##p = 0.001; unpaired t-test). c Analyses of transcript abundance of Nr1d1 at different circadian times (CT). Microglia were synchronized with dexamethasone (DEX, 100 nM) for 2 h and harvested every 4 h for RNA extraction and qPCR (n = 4). d Comparison of relative mRNA expression levels of Nr1d1 in acutely isolated microglia (n = 4 mice) (^##p = 0.0078) and peritoneal macrophages (n = 5 mice) (^##p = 0.0015) between WT and KI/+ mice (8-week-old, male). mRNA levels were normalized to 18s rRNA.e, f Representative bioluminescence recording of rhythmic Per2::Luc expression in primary microglia (e) and BMDM (f) from Per2::Luc;Psen2^+/+ or Per2::Luc;Psen2^N141I/+ mice. Cells were cultured and exposed to DEX (100 nM) for 2 h, followed by measurement of luciferase bioluminescence (each measurement for 1 min with 10-min interval) for 5 days. Circadian period and amplitude were analyzed in microglia (e; n = 3) (^#p = 0.0245) and BMDM (f; n = 5) (^#p = 0.02). Unpaired t-test for analysis. Data are mean ± SEM. ^#p < 0.05, ^##p < 0.01, and ^###p < 0.001 for the indicated comparisons. Source data are provided as a Source Data file.