Flow cytometry of the purified mouse BM Sca-1⁺ cells for surface markers analysis.

<p>(A) unlabeled Sca-1⁺ cells (control; A1), labeled cells showing 96.6% Sca-1⁺ (A2),1.7% c-kit⁺(A3), and 1.4% CD45⁺(A4) cell populations. (B) Immunostaining of cells for Sca-1 antigen (red = Sca-1; blue = DAPI; magnification = 400x). (C–E) Representative figures of <i>in vitro</i> characterization of BM Sca-1⁺ cells transfected for expression of the respective transgene. RT-PCR (C) and western blot (D) for HGF, IGF-1, SDF-1α, and VEGF transgene expression <i>in vitro</i>. In each case, non-transfected native Sca-1⁺ cells were used as a control. Densitometry showed significantly up-regulated expression of the respective growth factor gene and protein in the transfected Sca-1⁺ cells 48 hours after transfection. (E) Fluorescence images of the Sca-1⁺ cells immunostained for respective growth factor HGF (E1, red), IGF-1(E2, red), SDF-1α (E3, red), VEGF (E4, red). DAPI was used to visualize the nuclei (blue, magnification = 400x).