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. 2013 Jun 3;210(6):1137–1151. doi: 10.1084/jem.20122344

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© 2013 Roberts et al.

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Figure 4. — FAP⁺ cells of the bone marrow and hematopoiesis. (A) The cellularity of the femoral bone marrow from BAC transgenic mice and littermates was determined by hematoxylin and eosin staining after 3-d treatment with DTX. (B) Individual populations of hematopoietic cells from the femoral bone marrow of BAC transgenic mice and littermates were analyzed by flow cytometry after 3-d treatment of the mice with DTX. (C) 7 d after DTX administration, samples of peripheral blood from transgenic and littermate mice were assayed for hemoglobin, erythrocyte numbers, mean corpuscular hemoglobin, and mean corpuscular volume. (D) The proportions of FAP⁺ cells in enzyme-dissociated bone and bone marrow preparations from BAC transgenic mice and littermates were assessed by flow cytometry after 3-d treatment of the mice with DTX. (E) The mRNA levels of Cxcl12 and KitL in cells of the femoral bone marrow from BAC transgenic mice and littermates were measured by qRT-PCR after 3-d treatment of the mice with DTX. (F) The protein levels of Cxcl12 and KitL in lysates of bone marrow from BAC transgenic mice and littermates were measured by ELISA after 3-d treatment of the mice with DTX. (G) A cross-section of normal mouse femur was stained with antibodies to FAP, osteopontin, and osteocalcin and assessed by confocal microscopy. (H) Subsets of enzymatically dispersed cells from femoral bone and marrow were FACS purified, and the sorted populations were assessed by qRT-PCR for expression of FAP, Cxcl12, KitL, osteopontin, osteocalcin, and Lepr. Error bars represent SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are representative of at least two independent analyses (n ≥ 4; A, B, D, and E) or one confirmatory analysis (n = 6; F). Combined data are from three independent experiments (n ≥ 4; B).