Figure 4
Figure 4. BM reconstitution with uterine GFP+ blast: 12-month follow-up in vivo. Representative scatter plots illustrating percentages of GFP+ cells in the BM, blood, or spleen of the chimeric recipients (A; by FACS; N = 5). RT-PCR confirmed GFP mRNA expression in the chimeric BM and spleen (B). Control indicates wild-type BMCs (negative control); and positive, GFP+ transgenic BMCs (positive control). Recipient BMCs were cultured in MethoCult media. GFP+ colony-forming units were observed at day 7 (C). Representative micrographs illustrating coexpression of GFP and hematopoietic markers (CD16, CD13, CD45, and Mac-3) in BM isolates from the recipients (D, white arrows). DAPI indicates nuclear stain. Representative micrographs illustrating GFP expression (areas indicated in black boxes at right are enlarged at left, original magnification ×400) in formaldehyde-fixed tissue sections from multiple organs in the recipients (E, white arrows). Representative micrographs illustrating factor VIII and GFP expression (white arrows; colocalization and enlargement shown at bottom right) in the vasculature of frozen tissue sections from recipient somatic tissues (F).

BM reconstitution with uterine GFP+ blast: 12-month follow-up in vivo. Representative scatter plots illustrating percentages of GFP+ cells in the BM, blood, or spleen of the chimeric recipients (A; by FACS; N = 5). RT-PCR confirmed GFP mRNA expression in the chimeric BM and spleen (B). Control indicates wild-type BMCs (negative control); and positive, GFP+ transgenic BMCs (positive control). Recipient BMCs were cultured in MethoCult media. GFP+ colony-forming units were observed at day 7 (C). Representative micrographs illustrating coexpression of GFP and hematopoietic markers (CD16, CD13, CD45, and Mac-3) in BM isolates from the recipients (D, white arrows). DAPI indicates nuclear stain. Representative micrographs illustrating GFP expression (areas indicated in black boxes at right are enlarged at left, original magnification ×400) in formaldehyde-fixed tissue sections from multiple organs in the recipients (E, white arrows). Representative micrographs illustrating factor VIII and GFP expression (white arrows; colocalization and enlargement shown at bottom right) in the vasculature of frozen tissue sections from recipient somatic tissues (F).

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