The presumed atypical chemokine receptor CXCR7 signals through Gi/o proteins in primary rodent astrocytes and human glioma cells
Article first published online: 14 NOV 2011
Copyright © 2011 Wiley Periodicals, Inc.
Volume 60, Issue 3, pages 372–381, March 2012
How to Cite
Ödemis, V., Lipfert, J., Kraft, R., Hajek, P., Abraham, G., Hattermann, K., Mentlein, R. and Engele, J. (2012), The presumed atypical chemokine receptor CXCR7 signals through Gi/o proteins in primary rodent astrocytes and human glioma cells. Glia, 60: 372–381. doi: 10.1002/glia.22271
- Issue published online: 23 JAN 2012
- Article first published online: 14 NOV 2011
- Manuscript Accepted: 26 OCT 2011
- Manuscript Received: 12 JUL 2011
- DFG. Grant Number: KR 3408/2-1
Additional Supporting Information may be found in the online version of this article.
|GLIA_22271_sm_SuppFig1.doc||172K||Supporting Information Figure 1. SDF-1-induced chemotaxis of astrocytes is mediated by CXCR7. (A, B) Cortical rat astrocytes were transfected overnight with either selective CXCR7 siRNA (A) or selective CXCR4 siRNA (B) and analyzed for CXCR7 and CXCR4 expression, respectively, 72 h post-transfection. GAPDH served as loading control. (C) Analysis of astrocytes with RNAi-mediated inhibition of CXCR7 or CXCR4 expression for SDF-1-induced chemotaxis using a modified Boyden chamber. The number of spontaneously migrating cells present in transfection reagent-pretreated (control) cultures was set to 1. Transfection with CXCR7 siRNA, but not with CXCR4 siRNA prevented SDF-1-induced chemotaxis. Data represent mean + SD from 3 independent experiments. *p<0.05, **p<0.001; ANOVA with Student-Newman-Keuls; absence vs. presence of SDF-1.|
|GLIA_22271_sm_SuppFig2.doc||2473K||Supporting Information Figure 2. SDF-1-dependent activation of Akt and Erk in human glioma cells is sensitive to CCX771. Confluent cultures of human A764 glioma cells were maintained for 6 h with serum-free N2-medium and subsequently stimulated for 15 min with the indicated concentrations of SDF-1 in absence or presence of the CXCR7 antagonist, CCX771. Activation (phosphorylation) of Akt and Erk was assessed by Western blotting using phospho-specific antibodies. To control for protein loading, blot were additionally stained with antibodies recognizing Akt and Erk independent of their phosphorylation status. CCX771 prevented SDF-1-dependent activation of Akt and Erk, hence, confirming that activation of both signalling molecules occurs through CXCR7.|
|GLIA_22271_sm_SuppTab1.doc||255K||Supporting Information Table 1. Quantification of Western blot analysis. Integrated optical densities of immunoreactive protein bands were measured using the Gel-Pro Analyzer software. Data were normalized to the respective loading control. One way analysis of variance (ANOVA) followed by pairwise multiple comparison procedures (Student-Newman-Keuls method) was used for statistical analysis. Differences with p<0.05 were considered significant.|
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