American Society of Hematology

Figure 2

$Figure 2. Effect of Nanog overexpression on the differentiation of hematopoietic cells. Irradiated mice were reconstituted KSL cells transduced either by Gfp gene alone or Nanog-Iers-Gfp. Cells expressing the transgenes were distinguished by GFP expression. (A) Eight weeks later, the cells were harvested from various tissues of the recipients, and the proportion of GFP+ cells was measured by FACS. Compared with control recipients, the proportion of GFP+ cells in the peripheral blood, bone marrow, and spleen of the recipients remains at a low level. In contrast, nearly all cells in the thymus are GFP+. Bars represent a standard deviation. (B) Thymus atrophy in recipients of Nanog KSL cells (Nanog-thymus). Immunostaining by anti-Nanog antibody showed that Nanog-thymuses are occupied by cells expressing Nanog. The distinction of cortex and medulla, which is easily seen in the control thymus by hematoxylin staining, is difficult to discern in Nanog thymuses. The normal cortex of Nanog thymuses is replaced by lightly stained cells with larger cytoplasms. (C) Serial sections of thymuses were immunostained by antibodies to ER-TR4 and ER-TR5, which specify cortex and medulla, respectively. Thymocytes were also visualized by a mixture of antibodies to CD4 and CD8. Although the distinction between cortex and medulla is blurred as shown by HE staining, immunostaining by these antibodies demonstrate that each thymus compartment is preserved in the Nanog-thymus. In Nanog-thymuses, thymocytes expressing either CD4 or CD8 are present only in the medulla. Scale bar, 300 μm. (D) Integrin α4 and β1 expression of the control T cells expressing GFP and Nanog T cells. Almost all Nanog T cells express integrin α4 and integrinβ1, whereas only a small fraction of control thymocytes express it. FACS data of cells gated in GFP+ population is presented. (E) Integrin β7 expression in Nanog-thymus. The expression of integrin β7 was found only in the host T cells in the medulla of Nanog-thymus. However, Nanog expression is found in the cortex. Also in the control thymus, β7+ cells are localized in the medulla. Scale bar, 100 μm.$

Effect of Nanog overexpression on the differentiation of hematopoietic cells. Irradiated mice were reconstituted KSL cells transduced either by Gfp gene alone or Nanog-Iers-Gfp. Cells expressing the transgenes were distinguished by GFP expression. (A) Eight weeks later, the cells were harvested from various tissues of the recipients, and the proportion of GFP⁺ cells was measured by FACS. Compared with control recipients, the proportion of GFP⁺ cells in the peripheral blood, bone marrow, and spleen of the recipients remains at a low level. In contrast, nearly all cells in the thymus are GFP⁺. Bars represent a standard deviation. (B) Thymus atrophy in recipients of Nanog KSL cells (Nanog-thymus). Immunostaining by anti-Nanog antibody showed that Nanog-thymuses are occupied by cells expressing Nanog. The distinction of cortex and medulla, which is easily seen in the control thymus by hematoxylin staining, is difficult to discern in Nanog thymuses. The normal cortex of Nanog thymuses is replaced by lightly stained cells with larger cytoplasms. (C) Serial sections of thymuses were immunostained by antibodies to ER-TR4 and ER-TR5, which specify cortex and medulla, respectively. Thymocytes were also visualized by a mixture of antibodies to CD4 and CD8. Although the distinction between cortex and medulla is blurred as shown by HE staining, immunostaining by these antibodies demonstrate that each thymus compartment is preserved in the Nanog-thymus. In Nanog-thymuses, thymocytes expressing either CD4 or CD8 are present only in the medulla. Scale bar, 300 μm. (D) Integrin α4 and β1 expression of the control T cells expressing GFP and Nanog T cells. Almost all Nanog T cells express integrin α4 and integrinβ1, whereas only a small fraction of control thymocytes express it. FACS data of cells gated in GFP⁺ population is presented. (E) Integrin β7 expression in Nanog-thymus. The expression of integrin β7 was found only in the host T cells in the medulla of Nanog-thymus. However, Nanog expression is found in the cortex. Also in the control thymus, β7⁺ cells are localized in the medulla. Scale bar, 100 μm.

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