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. 2012 May 31;7(5):e38280. doi: 10.1371/journal.pone.0038280

Figure 2. Ectopic G0S2 expression reduces hematological reconstitution after BM transplantation.

Figure 2

BM cells were transduced with control MIGR1 or MIGR1-G0S2 (V5-tagged) retroviruses to study the effect of G0S2 on hematopoiesis. (A) Retroviral expression of G0S2 was measured by quantitative real-time PCR and immunoblotting (n = 3). (B) Transduced BM cells were used to study subcellular localization using anti-V5 (red), DAPI (blue) and anti-Nup98, COX IV, Calnexin or Rab5 (green) antibodies. The data represent three independent experiments. (C) The cell-cycle distribution of transduced BM cells was analyzed using nuclear staining with propidium iodine and flow cytometry (n = 3). (D) Transduced BM cells were transplanted into lethally irradiated mice and the frequency (CFU) and colony cell number were enumerated in methylcellulose culture (n = 3) after three months of hematologic reconstitution. (E) The contribution of donor-derived cells to myeloid and T cell populations in the peripheral blood was analyzed after transplantation by flow cytometry at different times post-transplant (n = 3–4). The data are representative of two independent experiments. *, P<0.05, **, P<0.01, and ***, P<0.001 (two-tailed Student's t-test).