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Blood, Vol. 113, Issue 24, 6085-6093, June 11, 2009 This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef CXCR7 heterodimerizes with CXCR4 and regulates CXCL12-mediated G protein signaling Blood Levoye et al. 113: 6085 Supplemental materials for: Levoye et al Files in this Data Supplement: Figure S1. Dose-dependent CXCL12-induced conformational changes in CXCR4 and CXCR7 homo- and heterodimers (JPG, 229 KB) - HEK-293T cells transfected with CXCR4-RLuc and CXCR4-YFP (A), CXCR7-Rluc and CXCR7-YFP (B), CXCR4-RLuc and CXCR7-YFP (C) were incubated with increasing concentrations of CXCL12 and BRET signal was measured as described in the Materials and Methods section. Results are expressed as percent of maximal ligand-induced BRET (set at 100%) and represent the means ± SEM of 3 independent experiments performed in duplicate. Figure S2. CXCR4-mediated G protein activation is not affected by the anti-CXCR7 blocking mAb (JPG, 115 KB) - 35S-GTPγS binding to membranes from HEK-293T cells expressing CXCR4 with or without CXCR7 was assessed upon increasing concentrations of CXCL12 in the presence or absence of the anti-CXCR7 9C4 mAb at 10 µg/ml. Results (means ± SEM) are from triplicate determinations. Figure S3. CXCR7 expression did not alter HIV infection of CD4-containing HEK-293T cells expressing CXCR4 and CCR5 (JPG, 186 KB) - (A and B) Cells described in the legend to Fig. 3D were infected or not for 2 h with R5-tropic BaL (30 ng of Gag p24/ml at the 1/100 virus dilution) (A), or X4-tropic NL4-3 (80 ng of Gag p24/ml at the 1/100 virus dilution) (B) HIV-1 particles carrying the firefly luc reporter gene in place of the HIV-1 nef gene. Fourty eight hours post-infection, cells were lysed and assessed for luciferase activity as reported.52 Results are expressed as relative luciferase units per microgram of cell lysate proteins. One of 2 independent experiments is shown. Figure S4. CXCL12-induced BRET changes between CXCR4 and Gαi1 correlate with G protein activation (JPG, 105 KB) - Dose-response of CXCL12-induced change in BRET signals was generated in cells expressing Gαi1-Rluc and CXCR4-YFP. BRET signals were measured as described in the Materials and Methods section. Results are expressed as percent of maximal CXCL12-induced change in BRET in each experiment (set at 100%) and are the means ± SEM of 3 independent experiments performed in duplicate. Figure S5. Kinetic analysis of receptors-Gαi1 interactions (JPG, 352 KB) - BRET kinetic analysis of the CXCL12-induced BRET increase in cells co-transfected with either Gαi1-Rluc or Gαi1-60-Rluc and CXCR4-YFP (A and C) or CXCR7-YFP (B and D). Repetitive signals were recorded before and immediately after CXCL12 (1 µM) injection (arrow) for 100 s. Data are representative of 3 independent experiments. This Article Abstract Full Text Services Email this article to a friend Alert me to new issues of the journal Rights and Permissions Citing Articles Citing Articles via CrossRef

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