Western blot analysis of cyclin D₁ in PB Inc⁻ CD34⁺ cells.

PB CD34⁺ cells immediately purified after density gradient separation (Inc⁻) were incubated (1 × 10⁵cells/mL) in a serum- and cytokine-free Stemα-A medium in the presence or absence of SDF-1 (0.5 ng/mL). After a 48-hour incubation, cells were harvested and processed for Western blot analysis of cyclin D₁ as indicated in “Materials and methods.” Biologic specificity of SDF-1 was demonstrated by using an anti–SDF-1 antibody (5 ng/mL). Results from a typical donor are presented.