Figure 7
Figure 7. Multilineage potential. (A) FACS blood plots of the white wild-type (WT), GFP donor, spleen microinjections (MI), GFP/white 2-color, and ablative liver and spleen HCT. (B) Representative images of GFP+ donor-derived immune cell types in the skin of 2-color, spleen microinjected HCT, and ablative spleen and liver HCT. (C) Phagocytic and nonphagocytic cells isolated from spleen or liver microinjected animals, and ablative HCT chimeras. (D) Mature macrophages isolated from microinjected and ablative HCT animals identified by F4/80 staining. (E) B cells isolated from microinjected and ablative HCT animals. Green particles are fluorescent E coli used in the phagocytic assay. (F) T cells isolated from microinjected and ablative HCT axolotls. (G) GFP immunostaining shows various levels of GFP expression across blood cells. Low levels of GFP expression can be detected in nucleated donor-derived erythrocytes (large white arrow). No GFP expression is detected in recipient erythrocytes (small white arrow). (H) PCR-based genotyping. Lane 1, GFP donor whole blood; lane 2, GFP donor red blood cell fraction; lane 3, spleen microinjected chimera; lane 4, irradiated and spleen transplanted chimera with no GVHD; lane 5, irradiated and transplanted chimera with GVHD; lane 6, GFP+ cephalic half and white tail chimera; lane 7, white wild-type axolotl; lane 8, water control. Generuler low range DNA ladder (25-700 bp) confirms the expected 173-bp amplified band of the GFP gene.

Multilineage potential. (A) FACS blood plots of the white wild-type (WT), GFP donor, spleen microinjections (MI), GFP/white 2-color, and ablative liver and spleen HCT. (B) Representative images of GFP+ donor-derived immune cell types in the skin of 2-color, spleen microinjected HCT, and ablative spleen and liver HCT. (C) Phagocytic and nonphagocytic cells isolated from spleen or liver microinjected animals, and ablative HCT chimeras. (D) Mature macrophages isolated from microinjected and ablative HCT animals identified by F4/80 staining. (E) B cells isolated from microinjected and ablative HCT animals. Green particles are fluorescent E coli used in the phagocytic assay. (F) T cells isolated from microinjected and ablative HCT axolotls. (G) GFP immunostaining shows various levels of GFP expression across blood cells. Low levels of GFP expression can be detected in nucleated donor-derived erythrocytes (large white arrow). No GFP expression is detected in recipient erythrocytes (small white arrow). (H) PCR-based genotyping. Lane 1, GFP donor whole blood; lane 2, GFP donor red blood cell fraction; lane 3, spleen microinjected chimera; lane 4, irradiated and spleen transplanted chimera with no GVHD; lane 5, irradiated and transplanted chimera with GVHD; lane 6, GFP+ cephalic half and white tail chimera; lane 7, white wild-type axolotl; lane 8, water control. Generuler low range DNA ladder (25-700 bp) confirms the expected 173-bp amplified band of the GFP gene.

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