Figure 6
Figure 6. Dynamic analysis of infiltration/expansion of EGFP cells in transplanted mice. Total BM or LSK cells were obtained from B6/EGFP mice expressing EGFP in all tissues and infused into B6 mice irradiated lethally, generating B6/tBM/EGFP or B6/LSK/EGFP mice. BM whole-mount specimens were examined at different time points by confocal microscopy (A,C) and isolated EGFP + hematopoietic cells were quantified by flow cytometry (B,D). (A) BM images of the B6/BM/EGFP mice on days 1, 7, and 28 illustrate: on day 1, massive destruction of extracellular matrix (red) and very few EGFP + cells (green) close to the bone edge; on day 7, remodeling of the extracellular matrix and increased number of EGFP+ cells, in islands beneath the bone edge; and by day 28, a complex network of extracellular matrix (red) and numerous EGFP+ cells evenly distributed. (B) Frequencies of EGFP-expressing cells in BM cells of the B6/BM/EGFP mice on days 1, 7, and 28 by flow cytometry. (C) BM images of the B6/LSK/EGFP mice on days 7, 14, and 28: at day 7, scarce clusters of EGFP+ cells appeared centrally located; at day 14, several clusters of EGFP+ cells were localized both centrally and peripherally, toward the bone edge; and at day 28, islands of EGFP+ cells were visible in the center of the tissue. (D) Frequencies of EGFP-expressing cells in BM cells of the B6/LSK/EGFP mice on days 7, 14, and 28, measured by flow cytometry. Nuclei were stained with DAPI. Scale bars are in micrometers. Related images appear in supplemental Video 7.

Dynamic analysis of infiltration/expansion of EGFP cells in transplanted mice. Total BM or LSK cells were obtained from B6/EGFP mice expressing EGFP in all tissues and infused into B6 mice irradiated lethally, generating B6/tBM/EGFP or B6/LSK/EGFP mice. BM whole-mount specimens were examined at different time points by confocal microscopy (A,C) and isolated EGFP + hematopoietic cells were quantified by flow cytometry (B,D). (A) BM images of the B6/BM/EGFP mice on days 1, 7, and 28 illustrate: on day 1, massive destruction of extracellular matrix (red) and very few EGFP + cells (green) close to the bone edge; on day 7, remodeling of the extracellular matrix and increased number of EGFP+ cells, in islands beneath the bone edge; and by day 28, a complex network of extracellular matrix (red) and numerous EGFP+ cells evenly distributed. (B) Frequencies of EGFP-expressing cells in BM cells of the B6/BM/EGFP mice on days 1, 7, and 28 by flow cytometry. (C) BM images of the B6/LSK/EGFP mice on days 7, 14, and 28: at day 7, scarce clusters of EGFP+ cells appeared centrally located; at day 14, several clusters of EGFP+ cells were localized both centrally and peripherally, toward the bone edge; and at day 28, islands of EGFP+ cells were visible in the center of the tissue. (D) Frequencies of EGFP-expressing cells in BM cells of the B6/LSK/EGFP mice on days 7, 14, and 28, measured by flow cytometry. Nuclei were stained with DAPI. Scale bars are in micrometers. Related images appear in supplemental Video 7.

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