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small molecules j113863  (Bio-Techne corporation)


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    Bio-Techne corporation small molecules j113863
    <t>J113863</t> and UCB35625 activate CCR2 and CCR5. A, structure of the small molecule enantiomers J113863 and UCB35625. B and C, competition binding assays performed on cells expressing CCR2 or CCR5 incubated, respectively, with 0.1 nm 125I-CCL2 or 125I-CCL4 as tracers and increasing concentrations of unlabeled CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm concentrations of unlabeled chemokine and specific binding in the absence of competitor (100%). D and E, chemotactic response of L1.2 cells stably expressing CCR2 or CCR5 in the presence of increasing concentrations of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). L1.2 cells expressing CCR5 were also co-stimulated with 50 μm J113863 and increasing concentrations of CCL4 (□). The data represent the mean values ± S.E. of three independent experiments.
    Small Molecules J113863, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/small molecules j113863/product/Bio-Techne corporation
    Average 93 stars, based on 25 article reviews
    small molecules j113863 - by Bioz Stars, 2026-02
    93/100 stars

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    1) Product Images from "Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 * "

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M116.757559

    J113863 and UCB35625 activate CCR2 and CCR5. A, structure of the small molecule enantiomers J113863 and UCB35625. B and C, competition binding assays performed on cells expressing CCR2 or CCR5 incubated, respectively, with 0.1 nm 125I-CCL2 or 125I-CCL4 as tracers and increasing concentrations of unlabeled CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm concentrations of unlabeled chemokine and specific binding in the absence of competitor (100%). D and E, chemotactic response of L1.2 cells stably expressing CCR2 or CCR5 in the presence of increasing concentrations of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). L1.2 cells expressing CCR5 were also co-stimulated with 50 μm J113863 and increasing concentrations of CCL4 (□). The data represent the mean values ± S.E. of three independent experiments.
    Figure Legend Snippet: J113863 and UCB35625 activate CCR2 and CCR5. A, structure of the small molecule enantiomers J113863 and UCB35625. B and C, competition binding assays performed on cells expressing CCR2 or CCR5 incubated, respectively, with 0.1 nm 125I-CCL2 or 125I-CCL4 as tracers and increasing concentrations of unlabeled CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm concentrations of unlabeled chemokine and specific binding in the absence of competitor (100%). D and E, chemotactic response of L1.2 cells stably expressing CCR2 or CCR5 in the presence of increasing concentrations of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). L1.2 cells expressing CCR5 were also co-stimulated with 50 μm J113863 and increasing concentrations of CCL4 (□). The data represent the mean values ± S.E. of three independent experiments.

    Techniques Used: Binding Assay, Expressing, Incubation, Stable Transfection

    Binding and signaling parameters of J113863 and UCB35625 Binding and functional parameters were measured on cells expressing CCR2 or CCR5. The pIC 50 and pEC 50 and Emax values were obtained from experiments as displayed in Figs. 1 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.
    Figure Legend Snippet: Binding and signaling parameters of J113863 and UCB35625 Binding and functional parameters were measured on cells expressing CCR2 or CCR5. The pIC 50 and pEC 50 and Emax values were obtained from experiments as displayed in Figs. 1 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.

    Techniques Used: Binding Assay, Functional Assay, Expressing

    A and B, UCB35625 and J113863 do not activate CCR1 and CCR3 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR1 (A) or CCR3 (B) were stimulated with CCL3 (♦) CCL11 (♢), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). C and D, UCB35625 and J113863 inhibited the activation of CCR1 and CCR3 triggered by chemokines. Cells stably expressing apoaequorin and CCR1 (C) or CCR3 (D) were incubated for 15 min with increasing concentrations of UCB35625 (●) or J113863 (○) before stimulation with 2 nm CCL3 or CCL11, and the luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 2 nm chemokine only (100%). E and F, UCB35625 and J113863 activate CCR2 and CCR5 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR2 (E) or CCR5 (F) were stimulated with CCL2 (■), CCL4 (▴), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). All the data represent the mean ± S.E. of three independent experiments.
    Figure Legend Snippet: A and B, UCB35625 and J113863 do not activate CCR1 and CCR3 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR1 (A) or CCR3 (B) were stimulated with CCL3 (♦) CCL11 (♢), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). C and D, UCB35625 and J113863 inhibited the activation of CCR1 and CCR3 triggered by chemokines. Cells stably expressing apoaequorin and CCR1 (C) or CCR3 (D) were incubated for 15 min with increasing concentrations of UCB35625 (●) or J113863 (○) before stimulation with 2 nm CCL3 or CCL11, and the luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 2 nm chemokine only (100%). E and F, UCB35625 and J113863 activate CCR2 and CCR5 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR2 (E) or CCR5 (F) were stimulated with CCL2 (■), CCL4 (▴), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). All the data represent the mean ± S.E. of three independent experiments.

    Techniques Used: Stable Transfection, Expressing, Activation Assay, Incubation

    Panel of G proteins activated by CCR2 upon stimulation by CCL2, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm CCL2, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01) and between ligands (###, p < 0.001) was assessed using Tukey's test.
    Figure Legend Snippet: Panel of G proteins activated by CCR2 upon stimulation by CCL2, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm CCL2, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01) and between ligands (###, p < 0.001) was assessed using Tukey's test.

    Techniques Used: Expressing

    Panel of G proteins activated by CCR5 upon stimulation by CCL4, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR5 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm of CCL4, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01; *, p < 0.1) and between ligands (##, p < 0.01; #, p < 0.1) was assessed using Tukey's test.
    Figure Legend Snippet: Panel of G proteins activated by CCR5 upon stimulation by CCL4, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR5 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm of CCL4, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01; *, p < 0.1) and between ligands (##, p < 0.01; #, p < 0.1) was assessed using Tukey's test.

    Techniques Used: Expressing

    Activation of Gi2 and Goa by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 (A and B) or CCR5 (C and D) and G protein biosensors and stimulated with increasing concentration of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments.
    Figure Legend Snippet: Activation of Gi2 and Goa by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 (A and B) or CCR5 (C and D) and G protein biosensors and stimulated with increasing concentration of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments.

    Techniques Used: Activation Assay, Concentration Assay

    Signaling parameters of J113863 and UCB35625 Functional parameters were measured on cells expressing CCR2 or CCR5. The pEC 50 and Emax values were obtained from experiments as displayed in Figs. 5 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.
    Figure Legend Snippet: Signaling parameters of J113863 and UCB35625 Functional parameters were measured on cells expressing CCR2 or CCR5. The pEC 50 and Emax values were obtained from experiments as displayed in Figs. 5 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.

    Techniques Used: Functional Assay, Expressing

    Recruitment of β-arrestin 2 by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells expressing either β-arrestin2-RLuc8 and CCR2-Venus (A and B) or β-arrestin2-RLuc8 and CCR5-Venus (C and D) and stimulated with CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). For kinetics, BRET signals were measured after the addition of 100 nm of chemokine or 100 μm J113863 or UCB35625. For dose-response curves, BRET was recorded 30 min after stimulation with various concentrations of ligands. Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.
    Figure Legend Snippet: Recruitment of β-arrestin 2 by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells expressing either β-arrestin2-RLuc8 and CCR2-Venus (A and B) or β-arrestin2-RLuc8 and CCR5-Venus (C and D) and stimulated with CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). For kinetics, BRET signals were measured after the addition of 100 nm of chemokine or 100 μm J113863 or UCB35625. For dose-response curves, BRET was recorded 30 min after stimulation with various concentrations of ligands. Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.

    Techniques Used: Expressing

    Transduction ratios log(τ/ K A ) of CCL4, UCB35625, and J113863 at CCR5 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL4 as the reference ligand. The RE of ligands toward each pathway, relative to CCL4, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.
    Figure Legend Snippet: Transduction ratios log(τ/ K A ) of CCL4, UCB35625, and J113863 at CCR5 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL4 as the reference ligand. The RE of ligands toward each pathway, relative to CCL4, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Techniques Used: Transduction

    Transduction ratios (log(τ/ K A )) of CCL2, UCB35625, and J113863 at CCR2 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL2 as the reference ligand. The RE of ligands toward each pathway, relative to CCL2, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.
    Figure Legend Snippet: Transduction ratios (log(τ/ K A )) of CCL2, UCB35625, and J113863 at CCR2 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL2 as the reference ligand. The RE of ligands toward each pathway, relative to CCL2, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Techniques Used: Transduction

    Bias factors for CCL4, UCB35625, and J113863 at CCR5 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL4, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.
    Figure Legend Snippet: Bias factors for CCL4, UCB35625, and J113863 at CCR5 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL4, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Techniques Used:

    Bias factors for CCL2, UCB35625, and J113863 at CCR2 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL2, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.
    Figure Legend Snippet: Bias factors for CCL2, UCB35625, and J113863 at CCR2 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL2, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Techniques Used:

    Functional response of CCR5 mutants in which aromatic residues in TM2 and TM3 were substituted by the corresponding amino acids of CCR2. Calcium mobilization measured in cells stably expressing apoaequorin and wild type CCR5 (black bar) or mutant forms of CCR5 bearing F2.39L, Y2.63S, F3.28L, F3.33H, or F3.36Y substitutions. Cells were stimulated either with CCL4 (A), UCB35625 (B), or J113863 (C), and luminescence was recorded for 30 s. The results were normalized for the basal luminescence in absence of agonist (0%) and the maximal response obtained with 50 μm APT (100%). Data represent the mean ± S.E. of three independent experiments. Statistical significance was assessed using Tukey's test (***, p < 0.001; *, p < 0.1).
    Figure Legend Snippet: Functional response of CCR5 mutants in which aromatic residues in TM2 and TM3 were substituted by the corresponding amino acids of CCR2. Calcium mobilization measured in cells stably expressing apoaequorin and wild type CCR5 (black bar) or mutant forms of CCR5 bearing F2.39L, Y2.63S, F3.28L, F3.33H, or F3.36Y substitutions. Cells were stimulated either with CCL4 (A), UCB35625 (B), or J113863 (C), and luminescence was recorded for 30 s. The results were normalized for the basal luminescence in absence of agonist (0%) and the maximal response obtained with 50 μm APT (100%). Data represent the mean ± S.E. of three independent experiments. Statistical significance was assessed using Tukey's test (***, p < 0.001; *, p < 0.1).

    Techniques Used: Functional Assay, Stable Transfection, Expressing, Mutagenesis

    F3.33H substitution does not impact significantly the binding of UCB35625 and J113863 to CCR5. Competition binding assays performed on cells expressing wild type CCR5 (A) or CCR5F3.33H (B) incubated with 0.1 nm 125I-CCL4 as tracer and increasing concentrations of unlabeled CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm of unlabeled chemokine and specific binding in the absence of competitor (100%).
    Figure Legend Snippet: F3.33H substitution does not impact significantly the binding of UCB35625 and J113863 to CCR5. Competition binding assays performed on cells expressing wild type CCR5 (A) or CCR5F3.33H (B) incubated with 0.1 nm 125I-CCL4 as tracer and increasing concentrations of unlabeled CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm of unlabeled chemokine and specific binding in the absence of competitor (100%).

    Techniques Used: Binding Assay, Expressing, Incubation

    Impact of F3. 33H substitution on cell chemotaxis, G protein activation, and arrestin recruitment. A and B, chemotactic response of L1.2 cells stably expressing CCR5 (●) or CCR5F3.33H (○) in the presence of increasing concentrations of J113863 (A) or UCB35625 (B). The data represent the mean values ± S.E. of three independent experiments. C and D, real-time measurement of BRET signal in HEK293T cells coexpressing the Gi2 biosensor and CCR5 (●) or CCR5F3.33H (○) and stimulated with increasing concentrations of J113863 (C) or UCB35625 (D). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments. E and F, real-time measurement of BRET signal in HEK293T cells expressing β-arrestin2-RLuc8 and CCR5-Venus (●) or β-arrestin2-RLuc8 and CCR5F3.33H-Venus (○) and stimulated with J113863 (E) or UCB35625 (F). Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus the receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.
    Figure Legend Snippet: Impact of F3. 33H substitution on cell chemotaxis, G protein activation, and arrestin recruitment. A and B, chemotactic response of L1.2 cells stably expressing CCR5 (●) or CCR5F3.33H (○) in the presence of increasing concentrations of J113863 (A) or UCB35625 (B). The data represent the mean values ± S.E. of three independent experiments. C and D, real-time measurement of BRET signal in HEK293T cells coexpressing the Gi2 biosensor and CCR5 (●) or CCR5F3.33H (○) and stimulated with increasing concentrations of J113863 (C) or UCB35625 (D). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments. E and F, real-time measurement of BRET signal in HEK293T cells expressing β-arrestin2-RLuc8 and CCR5-Venus (●) or β-arrestin2-RLuc8 and CCR5F3.33H-Venus (○) and stimulated with J113863 (E) or UCB35625 (F). Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus the receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.

    Techniques Used: Chemotaxis Assay, Activation Assay, Stable Transfection, Expressing



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    small molecules j113863  (Bio-Techne corporation)
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    Bio-Techne corporation small molecules j113863
    <t>J113863</t> and UCB35625 activate CCR2 and CCR5. A, structure of the small molecule enantiomers J113863 and UCB35625. B and C, competition binding assays performed on cells expressing CCR2 or CCR5 incubated, respectively, with 0.1 nm 125I-CCL2 or 125I-CCL4 as tracers and increasing concentrations of unlabeled CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm concentrations of unlabeled chemokine and specific binding in the absence of competitor (100%). D and E, chemotactic response of L1.2 cells stably expressing CCR2 or CCR5 in the presence of increasing concentrations of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). L1.2 cells expressing CCR5 were also co-stimulated with 50 μm J113863 and increasing concentrations of CCL4 (□). The data represent the mean values ± S.E. of three independent experiments.
    Small Molecules J113863, supplied by Bio-Techne corporation, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/small molecules j113863/product/Bio-Techne corporation
    Average 93 stars, based on 1 article reviews
    small molecules j113863 - by Bioz Stars, 2026-02
    93/100 stars
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    J113863 and UCB35625 activate CCR2 and CCR5. A, structure of the small molecule enantiomers J113863 and UCB35625. B and C, competition binding assays performed on cells expressing CCR2 or CCR5 incubated, respectively, with 0.1 nm 125I-CCL2 or 125I-CCL4 as tracers and increasing concentrations of unlabeled CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm concentrations of unlabeled chemokine and specific binding in the absence of competitor (100%). D and E, chemotactic response of L1.2 cells stably expressing CCR2 or CCR5 in the presence of increasing concentrations of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). L1.2 cells expressing CCR5 were also co-stimulated with 50 μm J113863 and increasing concentrations of CCL4 (□). The data represent the mean values ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: J113863 and UCB35625 activate CCR2 and CCR5. A, structure of the small molecule enantiomers J113863 and UCB35625. B and C, competition binding assays performed on cells expressing CCR2 or CCR5 incubated, respectively, with 0.1 nm 125I-CCL2 or 125I-CCL4 as tracers and increasing concentrations of unlabeled CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm concentrations of unlabeled chemokine and specific binding in the absence of competitor (100%). D and E, chemotactic response of L1.2 cells stably expressing CCR2 or CCR5 in the presence of increasing concentrations of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). L1.2 cells expressing CCR5 were also co-stimulated with 50 μm J113863 and increasing concentrations of CCL4 (□). The data represent the mean values ± S.E. of three independent experiments.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Binding Assay, Expressing, Incubation, Stable Transfection

    Binding and signaling parameters of J113863 and UCB35625 Binding and functional parameters were measured on cells expressing CCR2 or CCR5. The pIC 50 and pEC 50 and Emax values were obtained from experiments as displayed in Figs. 1 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Binding and signaling parameters of J113863 and UCB35625 Binding and functional parameters were measured on cells expressing CCR2 or CCR5. The pIC 50 and pEC 50 and Emax values were obtained from experiments as displayed in Figs. 1 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Binding Assay, Functional Assay, Expressing

    A and B, UCB35625 and J113863 do not activate CCR1 and CCR3 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR1 (A) or CCR3 (B) were stimulated with CCL3 (♦) CCL11 (♢), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). C and D, UCB35625 and J113863 inhibited the activation of CCR1 and CCR3 triggered by chemokines. Cells stably expressing apoaequorin and CCR1 (C) or CCR3 (D) were incubated for 15 min with increasing concentrations of UCB35625 (●) or J113863 (○) before stimulation with 2 nm CCL3 or CCL11, and the luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 2 nm chemokine only (100%). E and F, UCB35625 and J113863 activate CCR2 and CCR5 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR2 (E) or CCR5 (F) were stimulated with CCL2 (■), CCL4 (▴), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). All the data represent the mean ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: A and B, UCB35625 and J113863 do not activate CCR1 and CCR3 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR1 (A) or CCR3 (B) were stimulated with CCL3 (♦) CCL11 (♢), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). C and D, UCB35625 and J113863 inhibited the activation of CCR1 and CCR3 triggered by chemokines. Cells stably expressing apoaequorin and CCR1 (C) or CCR3 (D) were incubated for 15 min with increasing concentrations of UCB35625 (●) or J113863 (○) before stimulation with 2 nm CCL3 or CCL11, and the luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 2 nm chemokine only (100%). E and F, UCB35625 and J113863 activate CCR2 and CCR5 as measured by calcium mobilization. Cells stably expressing apoaequorin and CCR2 (E) or CCR5 (F) were stimulated with CCL2 (■), CCL4 (▴), UCB35625 (●), or J113863 (○), and luminescence was recorded for 30 s. The results were normalized for basal luminescence in the absence of agonist (0%), and maximal response was obtained with 50 μm ATP (100%). All the data represent the mean ± S.E. of three independent experiments.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Stable Transfection, Expressing, Activation Assay, Incubation

    Panel of G proteins activated by CCR2 upon stimulation by CCL2, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm CCL2, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01) and between ligands (###, p < 0.001) was assessed using Tukey's test.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Panel of G proteins activated by CCR2 upon stimulation by CCL2, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm CCL2, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01) and between ligands (###, p < 0.001) was assessed using Tukey's test.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Expressing

    Panel of G proteins activated by CCR5 upon stimulation by CCL4, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR5 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm of CCL4, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01; *, p < 0.1) and between ligands (##, p < 0.01; #, p < 0.1) was assessed using Tukey's test.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Panel of G proteins activated by CCR5 upon stimulation by CCL4, J113863, or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR5 and G protein biosensors (black bars) or G protein biosensors only (open bars) and stimulated for 1 min with 100 nm of CCL4, 1 μm TAK779, 100 μm UCB35625 (UCB), or 100 μm J113863 (J). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean values ± S.E. of six independent experiments. Statistical significance between cells expressing or not CCR2 (***, p < 0.001; **, p < 0.01; *, p < 0.1) and between ligands (##, p < 0.01; #, p < 0.1) was assessed using Tukey's test.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Expressing

    Activation of Gi2 and Goa by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 (A and B) or CCR5 (C and D) and G protein biosensors and stimulated with increasing concentration of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Activation of Gi2 and Goa by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells coexpressing CCR2 (A and B) or CCR5 (C and D) and G protein biosensors and stimulated with increasing concentration of CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Activation Assay, Concentration Assay

    Signaling parameters of J113863 and UCB35625 Functional parameters were measured on cells expressing CCR2 or CCR5. The pEC 50 and Emax values were obtained from experiments as displayed in Figs. 5 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Signaling parameters of J113863 and UCB35625 Functional parameters were measured on cells expressing CCR2 or CCR5. The pEC 50 and Emax values were obtained from experiments as displayed in Figs. 5 and . Values represent the mean ± S.E. of at least three independent experiments. NT, not tested; ND, not determined.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Functional Assay, Expressing

    Recruitment of β-arrestin 2 by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells expressing either β-arrestin2-RLuc8 and CCR2-Venus (A and B) or β-arrestin2-RLuc8 and CCR5-Venus (C and D) and stimulated with CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). For kinetics, BRET signals were measured after the addition of 100 nm of chemokine or 100 μm J113863 or UCB35625. For dose-response curves, BRET was recorded 30 min after stimulation with various concentrations of ligands. Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Recruitment of β-arrestin 2 by CCR2 and CCR5 upon stimulation by J113863 or UCB35625. Real-time measurement of BRET signal in HEK293T cells expressing either β-arrestin2-RLuc8 and CCR2-Venus (A and B) or β-arrestin2-RLuc8 and CCR5-Venus (C and D) and stimulated with CCL2 (▴), CCL4 (■), UCB35625 (●), or J113863 (○). For kinetics, BRET signals were measured after the addition of 100 nm of chemokine or 100 μm J113863 or UCB35625. For dose-response curves, BRET was recorded 30 min after stimulation with various concentrations of ligands. Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Expressing

    Transduction ratios log(τ/ K A ) of CCL4, UCB35625, and J113863 at CCR5 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL4 as the reference ligand. The RE of ligands toward each pathway, relative to CCL4, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Transduction ratios log(τ/ K A ) of CCL4, UCB35625, and J113863 at CCR5 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL4 as the reference ligand. The RE of ligands toward each pathway, relative to CCL4, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Transduction

    Transduction ratios (log(τ/ K A )) of CCL2, UCB35625, and J113863 at CCR2 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL2 as the reference ligand. The RE of ligands toward each pathway, relative to CCL2, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Transduction ratios (log(τ/ K A )) of CCL2, UCB35625, and J113863 at CCR2 Data were analyzed by non-linear regression using the operational model equation to determine the transduction ratios (log(τ/ K A )). Δlog(τ/ K A ) ratios were calculated from log(τ/ K A ) values considering CCL2 as the reference ligand. The RE of ligands toward each pathway, relative to CCL2, corresponds to the inverse logarithm of the Δlog(τ/ K A ) ratio. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Transduction

    Bias factors for CCL4, UCB35625, and J113863 at CCR5 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL4, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Bias factors for CCL4, UCB35625, and J113863 at CCR5 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL4, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques:

    Bias factors for CCL2, UCB35625, and J113863 at CCR2 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL2, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Bias factors for CCL2, UCB35625, and J113863 at CCR2 ΔΔlog(τ/ K A ) ratios between the pathways were calculated from the Δlog(τ/ K A ) values ( ). The ligand bias factors (BF), relative to CCL2, correspond to the inverse logarithm of the ΔΔlog(τ/ K A ) ratios. Data are the mean ± S.E. of 3–6 experiments. ND, not determined.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques:

    Functional response of CCR5 mutants in which aromatic residues in TM2 and TM3 were substituted by the corresponding amino acids of CCR2. Calcium mobilization measured in cells stably expressing apoaequorin and wild type CCR5 (black bar) or mutant forms of CCR5 bearing F2.39L, Y2.63S, F3.28L, F3.33H, or F3.36Y substitutions. Cells were stimulated either with CCL4 (A), UCB35625 (B), or J113863 (C), and luminescence was recorded for 30 s. The results were normalized for the basal luminescence in absence of agonist (0%) and the maximal response obtained with 50 μm APT (100%). Data represent the mean ± S.E. of three independent experiments. Statistical significance was assessed using Tukey's test (***, p < 0.001; *, p < 0.1).

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Functional response of CCR5 mutants in which aromatic residues in TM2 and TM3 were substituted by the corresponding amino acids of CCR2. Calcium mobilization measured in cells stably expressing apoaequorin and wild type CCR5 (black bar) or mutant forms of CCR5 bearing F2.39L, Y2.63S, F3.28L, F3.33H, or F3.36Y substitutions. Cells were stimulated either with CCL4 (A), UCB35625 (B), or J113863 (C), and luminescence was recorded for 30 s. The results were normalized for the basal luminescence in absence of agonist (0%) and the maximal response obtained with 50 μm APT (100%). Data represent the mean ± S.E. of three independent experiments. Statistical significance was assessed using Tukey's test (***, p < 0.001; *, p < 0.1).

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Functional Assay, Stable Transfection, Expressing, Mutagenesis

    F3.33H substitution does not impact significantly the binding of UCB35625 and J113863 to CCR5. Competition binding assays performed on cells expressing wild type CCR5 (A) or CCR5F3.33H (B) incubated with 0.1 nm 125I-CCL4 as tracer and increasing concentrations of unlabeled CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm of unlabeled chemokine and specific binding in the absence of competitor (100%).

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: F3.33H substitution does not impact significantly the binding of UCB35625 and J113863 to CCR5. Competition binding assays performed on cells expressing wild type CCR5 (A) or CCR5F3.33H (B) incubated with 0.1 nm 125I-CCL4 as tracer and increasing concentrations of unlabeled CCL4 (■), UCB35625 (●), or J113863 (○) as competitors. The data were normalized for nonspecific binding (0%) in the presence of 300 nm of unlabeled chemokine and specific binding in the absence of competitor (100%).

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Binding Assay, Expressing, Incubation

    Impact of F3. 33H substitution on cell chemotaxis, G protein activation, and arrestin recruitment. A and B, chemotactic response of L1.2 cells stably expressing CCR5 (●) or CCR5F3.33H (○) in the presence of increasing concentrations of J113863 (A) or UCB35625 (B). The data represent the mean values ± S.E. of three independent experiments. C and D, real-time measurement of BRET signal in HEK293T cells coexpressing the Gi2 biosensor and CCR5 (●) or CCR5F3.33H (○) and stimulated with increasing concentrations of J113863 (C) or UCB35625 (D). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments. E and F, real-time measurement of BRET signal in HEK293T cells expressing β-arrestin2-RLuc8 and CCR5-Venus (●) or β-arrestin2-RLuc8 and CCR5F3.33H-Venus (○) and stimulated with J113863 (E) or UCB35625 (F). Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus the receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: Partial Agonist and Biased Signaling Properties of the Synthetic Enantiomers J113863/UCB35625 at Chemokine Receptors CCR2 and CCR5 *

    doi: 10.1074/jbc.M116.757559

    Figure Lengend Snippet: Impact of F3. 33H substitution on cell chemotaxis, G protein activation, and arrestin recruitment. A and B, chemotactic response of L1.2 cells stably expressing CCR5 (●) or CCR5F3.33H (○) in the presence of increasing concentrations of J113863 (A) or UCB35625 (B). The data represent the mean values ± S.E. of three independent experiments. C and D, real-time measurement of BRET signal in HEK293T cells coexpressing the Gi2 biosensor and CCR5 (●) or CCR5F3.33H (○) and stimulated with increasing concentrations of J113863 (C) or UCB35625 (D). Results are expressed as the difference in BRET signal measured in the presence and absence of stimulation. Data represent the mean ± S.E. of three independent experiments. E and F, real-time measurement of BRET signal in HEK293T cells expressing β-arrestin2-RLuc8 and CCR5-Venus (●) or β-arrestin2-RLuc8 and CCR5F3.33H-Venus (○) and stimulated with J113863 (E) or UCB35625 (F). Results are expressed as net BRET, corresponding to the difference in BRET signal between cells expressing arrestin plus the receptor and cells expressing arrestin only. Data represent the mean ± S.E. of three independent experiments.

    Article Snippet: Chemokines and small molecules J113863 and UCB35625 were purchased from BioTechne.

    Techniques: Chemotaxis Assay, Activation Assay, Stable Transfection, Expressing