Figure 1
SDF-1α induces E-selectin expression in EPC.
(A) Levels of mSDF-1α in sera 4 h after wound bed injection of γmSDF-1α vs. PBS (n = 8 mice/group). (B) Measurement of levels of E-selectin in BM EPC by flow cytometry. BMC were incubated with Abs against CD34, KDR and E-selectin or isotype-matched control Abs. EPC (CD34+/KDR+) were gated (P2), counted and analyzed for levels of E-selectin (Q2-1). Bar graphs show % of EPC in BM mononuclear cells and relative levels of E-selectin in BM-EPC. Levels of E-selectin in EPC from mice injected with PBS were established as “1” and relative levels of E-selectin in EPC from mice injected with γmSDF-1α were normalized accordingly (n = 8 mice/group). (C) Measurement of % of EPC in peripheral blood MNC and relative levels of E-selectin in circulating EPC as described in (B) (n = 8 mice/group). (D) Immunoblotting analysis of E-selectin expression upon γhSDF-1α (100 ng/ml) stimulation in human EPC at various time points. β-actin served as a loading control. (E) Human EPC were stimulated with γhSDF-1α or BSA for 4 h and total RNA was extracted. Expression of extracellular matrix and adhesion molecules were analyzed using RT2-PCRArray. Expression of E-selectin was upregulated upon γmSDF-1α stimulation. Levels of mRNA in BSA-treated EPC were established as “1” and compared to those in γmSDF-1α-treated EPC. Experiments were repeated three times in (D) and (E). Data are analyzed by 2-tailed Student’s t-test and presented as mean ± SEM.
