SDF-1 overexpresses cyclin D1 and cyclin E in PB Inc−CD34+ cells.
PB CD34+ cells purified immediately after density gradient separation (Inc−) were incubated (1 × 105 cells/mL) in a serum- and cytokine-free Stemα-A medium in the presence or absence of SDF-1 (0.5 ng/mL). Freshly isolated cells (A) and cells harvested after a 48-hour incubation (B) were processed for intracellular detection of cyclin proteins as indicated in “Materials and methods.” The logarithm fluorescence of intracellular cyclin proteins is expressed in channel numbers. In panel A, histograms show results from staining with cyclin mAb (open histogram) and results from staining with irrelevant isotype-matched control antibody (solid histogram). Panel B shows histograms from cells stained with cyclin mAb after incubation in presence (open histogram) or absence (solid histogram) of SDF-1. Histograms from a typical donor are presented. The MFI (AU) of positive cells shown for each histogram are for one experiment representative of the 3 performed. Asterisk indicates P < .002 versus untreated cells.
