Figure 1. Nur77 is expressed in Ly6C⁻ monocytes.

(a) Relative mRNA expression of NR4A1 family members Nr4a1, Nr4a2, and Nr4a3 in wild-type bone marrow of FACS sorted Ly6C⁺, Ly6C⁻, MDP and CMP populations analyzed by qRT-PCR (n = 6 mice; expressed as a percentage of Ly6C⁺ monocyte transcript). (b) Nur77 protein expression in Ly6C⁺ and Ly6C⁻ monocyte populations from wild-type bone marrow, and CD11b⁻ non-myeloid cells measured by flow cytometric intracellular staining with a Nur77-specific antibody. For A and B, isolated monocyte populations were determined to be over 95% pure by Cytospin preps of sorted cells stained with HEMA3 dye. (c) Live F4/80⁺ CD11b⁺ monocytes from the peripheral blood of a Nur77-GFP transgenic reporter mouse and Nur77-GFP negative (GFP-) littermate control were assessed for Ly6C and GFP expression by flow cytometry. Left and center, representative gating strategy to identify Ly6C⁺ and Ly6C⁻ monocytes in the blood. Right, representative histogram of Ly6C⁺ and Ly6C⁻ monocytes, and cells from Nur77-GFP negative (GFP⁻) littermate control in the blood with a gate defining Nur77-GFP^high expression. (d) Circulating GFP^hi cells in Nur77-GFP reporter mice express other features of patrolling monocytes. Flow cytometry was performed on Nur77-GFP and control peripheral blood cells stained with antibodies indicated. Data from Nur77-GFP mice are representative of 4 experiments.