Figure 2.

Cold exposure modulates monocytes in the circulation

(A) Volcano plot showing the up- and downregulated transcripts by RNA sequencing of monocytes that were MACS (anti-PE) and FACS (CD115-PE+) sorted from blood of 2-week cold-exposed (10°C) mice compared to room-temperature mice.

(B) 15 most enriched Gene Ontology biological processes of monocytes as in (A). Genes were considered differentially regulated with p < 0.05. Shown are –log(p value).

(C) Blood immune cells of mice as in (A) visualized using UMAP and clustered using FlowSOM algorithm in R. Ly6C^hi monocytes (green), Ly6C^lo monocytes (yellow), neutrophils (brown), eosinophils (orange), and others. Heatmap shows median relative expression of all panel markers.

(D) Flow cytometry analysis of blood cells from mice as in (A). Percentage of Ly6C high (hi), intermediate (int), and low (lo) monocytes of total, single CD45⁺ cells.

(E) Percentage of blood MHCII⁺ cells of Ly6C^hi monocytes (left), MHCII⁺ Ly6C^hi monocytes of total MHCII⁺ cells (middle), and total MHCII⁺ cells of total, single CD45⁺ cells (right) of mice as in (A).

(F) Blood cell analysis after 1 week of daily i.p. injected beta 3 adrenoreceptor agonist CL316,243 or vector (PBS). Percentage of MHCII⁺ cells of Ly6C^hi monocytes (left), MHCII⁺ Ly6C^hi monocytes of total MHCII⁺ cells (middle), and total MHCII⁺ cells of CD45⁺ cells (right).

(D–F) Each dot represents one animal. Shown is mean ± SD. Significance was calculated using Student’s t test, ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001. Pool of three experiments (D and E).