Overexpression of Hmgb3 impairs myeloid and B-lymphocyte differentiation. (A) Diagram of Hmgb3-IRES-GFP retroviral vector and the control MSCV MGirL22Y IRES-GFP vector. (B) Mean CFU-C numbers per 1 × 105 cells plated for control IRES-GFP vector GFP– (n = 4; 223 colonies counted) and GFP+ (n = 3; 150 colonies counted) populations and for Hmgb3-IRES-GFP GFP– (n = 6; 424 colonies counted) and GFP+ (n = 6; 0 colonies counted) populations. (C) Representative FACS analysis of peripheral blood from mice who received transplants of either control IRES-GFP vector-transduced marrow (top: n = 8) or Hmgb3-IRES-GFP–transduced bone marrow (bottom: n = 10). Thymocytes, splenocytes, and bone marrow cells were analyzed 16 weeks after transplantation for presence of GFP in T cells, B cells, granulocytes/monocytes/macrophages, and erythroid cells, respectively. Quadrants were drawn based on isotype controls similar to those in Figure 3C. (D) Representative Southern blot analysis of genomic DNA isolated from bone marrow (M), thymus (T), and spleen (S) tissues from control GFP and Hmgb3-IRES-GFP mice 16-weeks after transplantation. 10 μg DNA were digested with EcoRI and loaded into each lane. Hybridization was performed with a probe that recognizes the GFP gene.
