GFP expression in bone marrow (BM) HSCs and MNCs. (A) Return of mobilized HSCs to the BM. Shown is the percentage of primitive (CD34⁺/CD45RA^–/CD90⁺) HSCs in total BM MNCs measured in BM samples at days 3 and 7. (“Day 0” is the day of HDAd injection.) (B) In vivo transduced primitive HSCs that returned to the BM. Shown is the percentage of GFP-positive CD34⁺/CD45RA^–/CD90⁺ HSCs in BM aspirates collected at days 3 and 7. (C) Shown are the data from panel B as fold increase, taking transduction with the HDAd5/35++GFP vector as “1.” (D) Percentage of GFP-positive CD34⁺/CD45RA^–/CD90⁺ HSCs relative to total BM MNCs. (E) Data extracted from panel D showing the effect of cxcr4 expression on the GFP⁺ HSCs in the BM (comparison of HDAd5/35++GFP vs HDAd5/35++GFP/cxcr4 and HDAd5/3+GFP vs HDAd5/3+GFP/cxcr4). (F) Representative GFP immunofluorescence on a BM CD34⁺ cytospin of the HDAd5/3+GFP/cxcr4–injected animal. CD34⁺ cells were isolated from a BM aspirate (after RBC lysis) by human CD34 magnetic-activated cell sorting, spun on a glass slide, and then stained with a GFP–fluorescein isothiocyanate antibody. GFP signals are green. Nuclei were stained with 4′,6-diamidino-2-phenylindole and appear in blue. Scale bar = 50 μm.