Journal: Cellular and Molecular Life Sciences

Article Title: Crosstalk between purinergic receptor P2Y 11 and chemokine receptor CXCR7 is regulated by CXCR4 in human macrophages

doi: 10.1007/s00018-024-05158-7

Figure Lengend Snippet: P2Y 11 and IL-1R cooperate to upregulate CXCR7 surface expression and CCL20 production in primary human M2 macrophages: potentiation by PDE4 inhibition. a M2 macrophages were treated for 24 h with ATPγS (20 µM) alone or in combination with IL-1ß (2 ng/ml) in the presence or absence of rolipram (10 µM). Flow cytometry was used to determine CXCR7 expression (light blue; isotype controls in red). Numbers are mean fluorescence intensities (MFIs) after subtraction of isotype control MFIs. b P2Y 11 receptor antagonist NF340 (20 µM) served to confirm that ATPγS-mediated changes were specific to P2Y 11 receptor activation. c Quantification of CXCR7 expression ( n = 3): mean values ± SD are shown. ** p < 0.01, *** p < 0.001, **** p < 0.0001. D) Quantification of CCL20 production after the same treatments that were used to upregulate CXCR7 expression ( n = 5). Mean values ± SD are shown. **** p < 0.0001

Article Snippet: The ATP analog ATPγS (Sigma Aldrich, St. Louis, MO, USA) and the suramin analog NF340 (Santa Cruz, Dallas, TX, USA) were utilized as P2Y 11 receptor agonist (20 μM) and antagonist (20 μM), respectively.

Techniques: Expressing, Inhibition, Flow Cytometry, Fluorescence, Control, Activation Assay

Journal: Cellular and Molecular Life Sciences

Article Title: Crosstalk between purinergic receptor P2Y 11 and chemokine receptor CXCR7 is regulated by CXCR4 in human macrophages

doi: 10.1007/s00018-024-05158-7

Figure Lengend Snippet: P2Y 11 activation is sufficient to induce high CXCR4 expression in primary human M2 macrophages: enhancement by PDE4 inhibition with no requirement for IL-1ß co-stimulation. a M2 macrophages were treated for 6 h with P2Y 11 receptor agonist ATPγS ± PDE4 inhibitor rolipram. Copy numbers of CXCR4 mRNA were determined using NanoString technology. P2Y 11 receptor antagonist NF340 was used to confirm that ATPγS-mediated changes were specific to P2Y 11 receptor activation. *** p < 0.001, **** p < 0.0001. b M2 macrophages were treated for 24 h with P2Y 11 receptor agonist ATPγS ± PDE4 inhibitor rolipram. Flow cytometry was used to determine CXCR4 expression (light blue; isotype controls in red). Numbers represent mean fluorescence intensities (MFIs) of the respective staining after subtraction of isotype control MFIs. c Quantification of CXCR4 expression ( n = 5). Mean values ± SD are shown. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The ATP analog ATPγS (Sigma Aldrich, St. Louis, MO, USA) and the suramin analog NF340 (Santa Cruz, Dallas, TX, USA) were utilized as P2Y 11 receptor agonist (20 μM) and antagonist (20 μM), respectively.

Techniques: Activation Assay, Expressing, Inhibition, Flow Cytometry, Fluorescence, Staining, Control

Journal: Cellular and Molecular Life Sciences

Article Title: Crosstalk between purinergic receptor P2Y 11 and chemokine receptor CXCR7 is regulated by CXCR4 in human macrophages

doi: 10.1007/s00018-024-05158-7

Figure Lengend Snippet: Synergistic cooperation between P2Y 11 and CXCR7 in primary human macrophages eliminates the need for IL-1ß co-stimulation and PDE4 inhibition. a M2 macrophages were treated for 24 h with P2Y 11 receptor agonist ATPγS and CXCR7 agonist TC14012, alone or in combination, and CCL20 was determined in culture supernatants ( n = 5). Mean values ± SD are shown. **** p < 0.0001. b Flow cytometry was used to determine CXCR7 and CXCR4 expression (light blue; isotype controls in red). Numbers represent mean fluorescence intensities (MFIs) of the respective staining after subtraction of isotype control MFIs. c Quantification of CXCR7 ( n = 5) and CXCR4 expression ( n = 3). Mean values ± SD are shown. *** p < 0.001, **** p < 0.0001. d M2 macrophages were treated for 24 h with P2Y 11 receptor agonist ATPγS plus IL-1ß in the presence or absence of either CXCR4 antagonist plerixafor or CXCR7 agonist TC14012. Pooled supernatants were analyzed for the presence of 20 CCL chemokines using RayBio technology. Plerixafor- or TC14012-mediated modulation of CCL chemokines that were present in the secretome and produced in a P2Y 11 receptor dependent manner (i.e. sensitive to inhibition with NF340) is shown (see also Fig. )

Article Snippet: The ATP analog ATPγS (Sigma Aldrich, St. Louis, MO, USA) and the suramin analog NF340 (Santa Cruz, Dallas, TX, USA) were utilized as P2Y 11 receptor agonist (20 μM) and antagonist (20 μM), respectively.

Techniques: Inhibition, Flow Cytometry, Expressing, Fluorescence, Staining, Control, Produced