Figure 6. Pregnancy increases Cyp27a1 expression and 27HC levels in primitive hematopoietic stem and progenitor cells.

(A and B) Cyp27a1 deficiency did not affect the numbers of hematopoietic stem and progenitor cells in the bone marrow (femurs and tibias; A) or spleen (B) of mice that had been repeatedly bled (a total of 5–6 mice/treatment from 3 independent experiments). (C and D) Cyp27a1 deficiency did not affect the numbers of hematopoietic stem and progenitor cells in the bone marrow (femurs and tibias; C) or spleen (D) of mice treated with G-CSF for 6 days (a total of 3 mice/treatment from 2 independent experiments). (E, G, and H) Cyp27a1 (E) or Cxcr4 (G and H) transcript levels were quantified by qRT-PCR in the indicated bone marrow cell populations (a total of 3–5 mice from 3–4 independent experiments). Expression values were normalized to beta-Actin and presented relative to those in control WBM cells. (F) Intracellular 27HC levels in 10⁷ lineage-depleted bone marrow cells from non-pregnant and pregnant WT mice (a total of 5 mice/treatment from 4 independent experiments). Statistical significance was assessed using 1-way ANOVA with Šídák’s multiple comparisons tests, with the exception of E, F, and H, where we used 2-tailed unpaired Student’s t tests using the FDR method to correct for multiple comparisons (*P < 0.05, ^‡P < 0.01, ^#P < 0.001). All data represent mean ± SD.