Figure 5. Accumulation of bone marrow monocytes in mice housed at thermoneutrality.

A) C57bl/6 mice were housed at 22°C or 30°C environment for seven days, then assayed for circulating monocytes (Ly6C⁺ and Ly6C⁻). B) C57bl/6 mice housed at indicated temperatures were treated with BrdU i.p. for 24hrs, then assayed for incorporation of BrdU in Ly6C⁺ monocytes to measure bone marrow egress C) Snapshot from 2-photon imaging approach of bone marrow in Cx3cr1^gfp reporter mice crossed with Cxcl12^dsred to examine monocytes within the bone marrow. D) Measurement of Cx3cr1^gfp expression per frame, Cxcl12^dsred expression per frame, and ratio of GFP/Dsred in bone marrow from animals housed at indicated temperature. E) Flow cytometeric analysis of Cx3cr1^gfp/+ bone marrow total cellularity and GFP⁺ monocytes housed at indicated ambient temperature. F) CCR2 MFI for bone marrow monocytes from Cx3cr1^gfp mice housed at 22°C or 30°C. (A-B, E-F) Data are representative experiments from three independent experiments with n≥5 per group. C) Image is a representative image of greater than 5 independent experiments. D) Data are combined from 5 independent experiments for each group. Statistical analysis was performed by unpaired student T-test, * p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001.