Human CXCR4 Antibody

Detection of CXCR4 by Flow Cytometry

Effect of hypoxia on CX3CR1 and CXCR4 expression by early culture of MSCs.(A) The MSCs were cultured for 4–20 h in normoxic (Nor) or hypoxic (Hyp) conditions, as indicated. Total RNA was analyzed by RT-PCR for CX3CR1, CX3CR4, and b-actin mRNA expression. (B) Total RNA from MSCs cultured for 20 h in the presence of increasing concentrations of DFX was analyzed by RT-PCR for CX3CR1 and CXCR4 mRNA expression. (C) Quantitaive RT-PCR for CXCR4 and CX3CR1 mRNA expression at 20 h of treatment. The vertical axis represents the relative ratio of indicated condition to normoxic condition without the addition of DFX.. (D) Surface expression of CXCR4 and CX3CR1 were determined by flow cytometry using a mouse mAb anti-CXCR4 and a rat mAb anti-CX3CR1, respectively (continuous line). Irrelevant antibody is indicated by the dotted line. The results are representative of three independent experiments. (E) Hypoxia and DFX-treated MSCs were cultured for 24 h in the indicated conditions and analyzed by western blot for HIF-1 alpha, CXCR4, CX3CR1, and actin protein levels. (F) HIF-1 alpha binding to the CX3CR1 promoter. The MSCs were cultured for 4 h in normoxic or hypoxic condition. ChIP was performed with or without rabbit antibody specific for HIF-1 alpha. There is no difference between PCR of the CX3CR1 promoter using input chromatin for normoxic and hypoxic cells. However, a band was noted for hypoxic cells but not for normoxic cells, when PCR of the CX3CR1 promoter was performed using immunoprecipitation with HIF-1 alpha antibody. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/17476338), licensed under a CC-BY license. Not internally tested by R&D Systems.