American Society of Hematology

Figure 5.

The D2-type receptor agonist 7-OH-DPAT promotes BM transplantation efficiency. Representative confocal images showing synaptophysin (A) and Th. The P value was calculated using the Student t test. (B) immunosignal in control (Ctrl) and irradiated BM at 7 days after 12 Gy irradiation. Quantification of the synaptophysin+ and Th+ puncta per length nerve. Ctrl = 16 slices from 3 mice; 12 Gy = 16 slices from 3 mice. (C) ELISA of dopamine levels in flushed BM extracellular fluid in control (ctrl) femur (n = 7) or 7 days after lethal irradiation (12 Gy) (n = 8). The P value was calculated using the Student t test. (D) Transplantation of Lin− cells from Vav1-Cre Rosa26-mTmG mice into 7-OH-DPAT–treated (short regimen, as indicated) or vehicle control mice after lethal irradiation. (E) Overview of bone sections from 7-OH-DPAT–treated or control mice after lethal irradiation and transplantation of GFP-labeled donor cells. (F) Representative confocal images showing GFP+ donor cells (green) and EdU incorporation (red) in sections from vehicle- and 7-OH-DPAT–treated long bone at 5 days after lethal irradiation and transplantation. (G) Representative FACS plot of GFP+ cells in irradiated and transplanted mice treated with vehicle or 7-OH-DPAT. (H) Quantification of the percentages of GFP+ cells and donor-derived CD45+ cells in BM by FACS (vehicle = 9; 7-OH-DPAT = 8). Quantification of EdU+ cells by imaging (vehicle = 5; 7-OH-DPAT = 5). The P value was calculated using the Student t test. (I) Schematic depiction of the transplantation of Lin− cells into 7-OH-DPAT–treated or vehicle control recipients (long treatment regimen) at day 21 after lethal irradiation. (J) Confocal images showing Ki67+ signal in bone sections from recipients treated with vehicle or 7-OH-DPAT. (K) Quantification of the numbers of BMNCs and CD45+ cells and the percentages of LSK cells (vehicle = 7; 7-OH-DPAT = 8). The P value was calculated using the Student t test. Quantification of Ki67+ cell number in each image field in panel J (vehicle = 4; 7-OH-DPAT = 5). (L) Representative images (dish at day 8 after seeding) (left panels) and quantification of CFU from 200 HSCs isolated from WT mice (right panel). 7-OH-DPAT or vehicle was added to MethoCult medium (vehicle = 12; 7-OH-DPAT = 12). The P value was calculated using the Student t test. 1h, 1 hour; D, day; SSC, scatter.

The D₂-type receptor agonist 7-OH-DPAT promotes BM transplantation efficiency. Representative confocal images showing synaptophysin (A) and Th. The P value was calculated using the Student t test. (B) immunosignal in control (Ctrl) and irradiated BM at 7 days after 12 Gy irradiation. Quantification of the synaptophysin⁺ and Th⁺ puncta per length nerve. Ctrl = 16 slices from 3 mice; 12 Gy = 16 slices from 3 mice. (C) ELISA of dopamine levels in flushed BM extracellular fluid in control (ctrl) femur (n = 7) or 7 days after lethal irradiation (12 Gy) (n = 8). The P value was calculated using the Student t test. (D) Transplantation of Lin⁻ cells from Vav1-Cre Rosa26-mTmG mice into 7-OH-DPAT–treated (short regimen, as indicated) or vehicle control mice after lethal irradiation. (E) Overview of bone sections from 7-OH-DPAT–treated or control mice after lethal irradiation and transplantation of GFP-labeled donor cells. (F) Representative confocal images showing GFP⁺ donor cells (green) and EdU incorporation (red) in sections from vehicle- and 7-OH-DPAT–treated long bone at 5 days after lethal irradiation and transplantation. (G) Representative FACS plot of GFP⁺ cells in irradiated and transplanted mice treated with vehicle or 7-OH-DPAT. (H) Quantification of the percentages of GFP⁺ cells and donor-derived CD45⁺ cells in BM by FACS (vehicle = 9; 7-OH-DPAT = 8). Quantification of EdU⁺ cells by imaging (vehicle = 5; 7-OH-DPAT = 5). The P value was calculated using the Student t test. (I) Schematic depiction of the transplantation of Lin⁻ cells into 7-OH-DPAT–treated or vehicle control recipients (long treatment regimen) at day 21 after lethal irradiation. (J) Confocal images showing Ki67⁺ signal in bone sections from recipients treated with vehicle or 7-OH-DPAT. (K) Quantification of the numbers of BMNCs and CD45⁺ cells and the percentages of LSK cells (vehicle = 7; 7-OH-DPAT = 8). The P value was calculated using the Student t test. Quantification of Ki67⁺ cell number in each image field in panel J (vehicle = 4; 7-OH-DPAT = 5). (L) Representative images (dish at day 8 after seeding) (left panels) and quantification of CFU from 200 HSCs isolated from WT mice (right panel). 7-OH-DPAT or vehicle was added to MethoCult medium (vehicle = 12; 7-OH-DPAT = 12). The P value was calculated using the Student t test. 1h, 1 hour; D, day; SSC, scatter.

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