Fig. 1. Nr4a1-GFP^high monocytes patrol the vasculature, and interact with tumor in the lung.

(A) Quantification of Nr4a1-GFP^high PMo per μl of blood volume in lung (Untreated), 4 hrs or 24 hrs after IV LLC-RFP transfer (n=5 mice per group). (B) Quantification of Nr4a1-GFP^high monocyte movement in lung before (Untreated), 4 hrs or 24 hrs after LLC-RFP tumor injection. Monocyte tracks transposed to a common origin from a representative 20 min movie (left, scale bar=100 μm), and quantification of median speed of monocytes (right, combined speed data from analysis of 3 separate mice; *=p<0.001 lower than untreated; **=p<0.001 than 4hr tumor). (C) Representative gating of Nr4a1-GFP^highCD11b⁺ cells from all Live CD45⁺CD11c^low cells 24 hrs after IV LLC-RFP transfer. (D) Representative confocal image of Nr4a1-GFP^high monocytes (Green) interacting with LLC-RFP cells (Red) in the lung 7 days after IV LLC-RFP transfer. Immune cells in the vasculature were labeled with IV injected anti-CD45 antibody (Blue). (E) Quantification of free (>100 μm from tumor site) and tumor-associated (<50 μm from tumor site) Nr4a1-GFP^high monocytes in the lung at various time points after tumor injection (combined analysis of 5 mice per group; p<0.01 for each tumor-associated area compared to free tumor area for each time point).