GPR84 knockdown blocks dissemination of leukemic cells to the spleen and compromises leukemia reconstitution. (A) Kaplan-Meier survival curves for mice injected with 1 × 10⁶ Scr or GPR84 KD#1 KLSMLL GFP⁺ pre-LSCs. (B) Percentage of GFP⁺ leukemic cells in BM and spleen. NS, not significant (P > .05). (C) Spleen weights of primary AML mice. (D-E) Flow cytometric analysis of GPR84 and β-catenin protein levels in GFP⁺ leukemic cells from primary recipients. (F-G) Kaplan-Meier survival curves for mice that received 1 × 10⁵ GFP⁺ leukemic cells from primary recipients in secondary transplantation assays, or that received the indicated number (10²-10⁴) of GFP⁺ leukemic cells for in vivo limiting dilution assessment. Calculated LSC frequencies in GFP⁺ Scr and GPR84 KD#1 leukemic cells are 1 in 433 and 1 in 1254, respectively. (H-I) Representative histograms illustrating in vivo BrdU incorporation cell proliferation assays of GFP⁺ leukemic cells from the BM of secondary recipients 10 days after transplantation. (J) Bar graphs representing quantitative analysis of the data obtained from in vivo BrdU incorporation assays (indicative of 6 independent experiments).