cx 3 cr1 inhibitor jms 17 2  (MedChemExpress)

Journal: npj Viruses

Article Title: Respiratory syncytial virus glycoprotein G impedes CX 3 CR1-activation by CX 3 CL1 and monocyte function

doi: 10.1038/s44298-024-00075-9

Figure Lengend Snippet: A Dose-response curve showing CX 3 CR1 activation in Chem-4 cells stimulated with increasing concentrations of CX 3 CL1 (0, 1, 5, 10, 50, 100, 500 nM). Cells pretreated with 10 µM JMS-17-2 inhibitor served as controls, with subsequent stimulation using the same concentrations of CX 3 CL1. Fluorescence intensity, indicative of calcium mobilization, was measured over 180 s. Data represent mean values ± SD ( n = 6 from three independent experiments). B CX 3 CR1 activation in Chem-4 cells stimulated with recombinant RSV sG (WT or CX 3 C Mut ) at concentrations equivalent to those in ( A ). The red dotted line indicates the mean relative fluorescence units (RFU) in response to 500 nM CX 3 CL1 (as shown in ( A )). Data represent mean values ± SD (n = 3 from three independent experiments). p values were determined by Welsh t-tests compared as indicated. C Receptor competition assay with 150 nM CX 3 CL1 and varying molar ratios of RSV sG WT or RSV sG CX 3 C Mut (5:1, 1:1, 1:2, 1:10, 1:20 CX 3 CL1) compared to untreated cells. Fluorescence intensity was measured to assess calcium flux and receptor activation. Mean values ± SD are shown ( n = 6 from three independent experiments). p values were determined by Dunnett’s One-way ANOVA compared to untreated cells. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: To assess the assay specificity, Chem-4 cells were additionally treated with 10 μM of the CX 3 CR1 inhibitor JMS-17-2 (MedChemExpress) or the corresponding concentration of DMSO for the generation of the dose response curve.

Techniques: Activation Assay, Fluorescence, Recombinant, Competitive Binding Assay

Journal: npj Viruses

Article Title: Respiratory syncytial virus glycoprotein G impedes CX 3 CR1-activation by CX 3 CL1 and monocyte function

doi: 10.1038/s44298-024-00075-9

Figure Lengend Snippet: A Flow cytometry analysis of CX 3 CR1 expression on THP-1 cells compared to HEK293 cells (negative control). B Quantification of THP-1 cell migration in response to 300 nM rBSA, 50 nM, 150 nM, and 300 nM RSV A rsG WT, 50 nM, 150 nM, and 300 nM RSV A rsG CX 3 C Mut , 50 nM, 150 nM, and 300 nM CX 3 CL1. Data represent mean values ± SD ( n = 6 from three independent experiments). Statistical significance was determined using One-Way ANOVA and Kruskal-Wallis tests compared to untreated cells. C THP-1 cell migration in response to 150 nM CX 3 CL1 in cells pretreated with 10 μM AZD8797, or equimolar ratios of rBSA, RSV A rsG WT, RSV A rsG CX 3 C Mut . Data represent mean values ± SD ( n = 6 from three independent experiments). Statistical significance was determined using One-Way ANOVA and Kruskal-Wallis tests compared to 150 nM CX 3 CL1 stimulation on untreated THP-1 cells. D Migration of THP-1 cells induced by supernatants from A549 cells treated with 300 nM or 150 nM RSV A rsG WT, 300 nM or 150 nM RSV A rsG CX 3 C Mut , 150 nM rBSA, or infected with RSV A 0594 at MOI 0.1, 1, or 3. Data represent mean values ± SD ( n = 6 from three independent experiments). Statistical significance was determined using One-Way ANOVA and Kruskal-Wallis tests compared to untreated cells. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: To assess the assay specificity, Chem-4 cells were additionally treated with 10 μM of the CX 3 CR1 inhibitor JMS-17-2 (MedChemExpress) or the corresponding concentration of DMSO for the generation of the dose response curve.

Techniques: Flow Cytometry, Expressing, Negative Control, Migration, Infection

Journal: npj Viruses

Article Title: Respiratory syncytial virus glycoprotein G impedes CX 3 CR1-activation by CX 3 CL1 and monocyte function

doi: 10.1038/s44298-024-00075-9

Figure Lengend Snippet: A Immunofluorescence staining of A549 cells for CX 3 CL1 expression after 12-h treatment with 30 µg/mL LTA, 10 µg/mL LPS, 150 nM RSV A sG WT, 150 nM RSV A sG CX 3 C Mut , and infection with RSV A 0594 at MOI 1. CX 3 CL1-FITC antibody staining shows the level of CX 3 CL1 expression under each condition. B Quantification of monocyte attachment to A549 cells pretreated with 10 µg/mL LPS and subsequently exposed 7.5 µM AZD8797, 150 nM or 300 nM RSV A sG WT, 150 nM or 300 nM RSV A sG CX 3 C Mut , 150 nM or 300 nM rBSA. Monocyte attachment was assessed by counting calcein-positive THP-1 cells using an ImageJ automated cell counting macro adopted from ref. . Data represent mean values ± SD ( n = 27 from three independent experiments). Statistical significance was determined using Dunnett’s One-way ANOVA compared to untreated cells. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: To assess the assay specificity, Chem-4 cells were additionally treated with 10 μM of the CX 3 CR1 inhibitor JMS-17-2 (MedChemExpress) or the corresponding concentration of DMSO for the generation of the dose response curve.

Techniques: Immunofluorescence, Staining, Expressing, Infection, Cell Counting