Mobilization efficacy and safety. (A) Left panel: number of LSK cells perμl blood at 2 hours after WU-106 + AMD3100 injection or 45 minutes after AMD3100 injection in the G-CSF + AMD3100 regimen (before the HDAd injection). LSK cells were measured by flow cytometry. Right panel: CFU were measured by methylcellulose-based colony assay with blood samples drawn at the above-listed time points. PBMCs from 5 μL mobilized peripheral blood were plated for the CFU assay. Total colonies were counted 11 days later. Each dot represents 1 mouse. (B) White blood cells (WBCs) in blood measured by hemocytometer after lysing RBCs. (C) Blood cell analysis. Hematological parameters measured by Hemavet. The platelet (PLT) counts are shown in the right panel. (D) Percentage of activated (CD11b⁺/Ly6G⁺) neutrophils at the peak of mobilization. (E) Serum cytokine levels 6 hours after HDAd injection were measured using a cytometric bead array. IL-6 levels are shown. (F) Serum CD62p (P-selectin) concentrations measured by enzyme-linked immunosorbent assay at the pak of mobilization (right before HDAd injection) “0 hours” and 6 hours after HDAd injection (“6 hours”). Studies in panels A-D were carried out in mice without HDAd injection at the peak of mobilization. Serum cytokine levels and platelets (E-F) were measured 6 hours after HDAd injection. For comparisons of multiple groups, a 2-way analysis of variance (ANOVA) with Bonferroni posttest for multiple comparisons was used. Statistical analysis was performed using GraphPad Prism version 10.0.3. ∗∗∗P ≤ .001, ∗∗P ≤ .01, ∗P ≤ .05. EO, eosinophils; Hb, hemoglobin; HCT, hematocrit; LY, lymphocytes; MCH, mean corpuscular Hb; MCHC, MCH concentration; MCV, mean corpuscular volume; MO, monocytes; NE, neutrophils.