Effects of iron deficiency on MPN phenotype. (A) Schematic drawing of the experimental setup for the BM transplantations and induction of iron deficiency. (B) Time course of body weight (n = 6-8 mice per group). (C) Complete blood counts (n = 6-8 mice per group) are shown at the indicated times. (D) The numbers of megakaryocytes (Mks) per 10 high-power fields (HPFs) counted in sternum sections of mice at original magnification ×400 are shown (left); n = 3 mice per genotype and treatment group. Thrombopoietin (Tpo) concentration in BM lavage (1 femur and 1 tibia) is shown (right). Dotted line indicates the limit of detection of the assay. (E) Iron parameters and regulatory cytokines. Group size was n = 4 to 8 mice per genotype and treatment group. The data presented in panels D and E were obtained at terminal workup, 8 weeks after start of treatment for Ki mice, at 12 weeks for E12 mice, and at 14 weeks for WT mice. Note that logarithmic scales are used for iron regulatory cytokines (Epo, erythroferrone, and hepcidin). All data are presented as mean ± standard error of the mean. Two-way ANOVA with subsequent Tukey test or Dunnett posttest was used for multiple-group comparisons. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, ∗∗∗∗P < .0001. CD, control diet; LD, low-iron diet; LD+Phl, low-iron diet plus phlebotomy.