ep2  (Alomone Labs)


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    Structured Review

    Alomone Labs ep2
    Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of <t>EP2</t> agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values
    Ep2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ep2/product/Alomone Labs
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    ep2 - by Bioz Stars, 2022-11
    91/100 stars

    Images

    1) Product Images from "Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study"

    Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

    Journal: Digestive Diseases and Sciences

    doi: 10.1007/s10620-019-05623-5

    Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of EP2 agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values
    Figure Legend Snippet: Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of EP2 agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values

    Techniques Used: Chemotaxis Assay, Incubation

    Immunohistochemistry shows strong DP1 staining in eosinophils ( a , arrows ), while epithelial cells in esophageal mucosal biopsies are practically devoid of staining. EP4 ( b ) and EP2 ( c ) immunostaining is prominent in eosinophils ( arrows ) but also visible in epithelial cells ( arrowheads ). 3-3′-Diaminobenzidine (DAB) was used as visualization substrate for DP1 and EP4 staining (brown precipitates), while for EP2, ImmPact™ NovaRED substrate was used (red precipitate). Calibration bar: 50 µm
    Figure Legend Snippet: Immunohistochemistry shows strong DP1 staining in eosinophils ( a , arrows ), while epithelial cells in esophageal mucosal biopsies are practically devoid of staining. EP4 ( b ) and EP2 ( c ) immunostaining is prominent in eosinophils ( arrows ) but also visible in epithelial cells ( arrowheads ). 3-3′-Diaminobenzidine (DAB) was used as visualization substrate for DP1 and EP4 staining (brown precipitates), while for EP2, ImmPact™ NovaRED substrate was used (red precipitate). Calibration bar: 50 µm

    Techniques Used: Immunohistochemistry, Staining, Immunostaining

    Flow cytometric evaluation of EP (EP1–4) and DP (DP1, DP2) receptor expression in human blood eosinophils. Changes in receptor expression were recorded as mean fluorescence intensity and data are expressed as fold increase in fluorescence over isotype control. Expression levels of EP2 and EP4 are significantly lower in patients with eosinophilic esophagitis (EoE) compared to healthy control subjects (control). Expression levels of EP1, EP3, DP1, and DP2 do not differ between EoE and controls. Values are mean ± SEM; Student’s t test; n = 6–7. * p
    Figure Legend Snippet: Flow cytometric evaluation of EP (EP1–4) and DP (DP1, DP2) receptor expression in human blood eosinophils. Changes in receptor expression were recorded as mean fluorescence intensity and data are expressed as fold increase in fluorescence over isotype control. Expression levels of EP2 and EP4 are significantly lower in patients with eosinophilic esophagitis (EoE) compared to healthy control subjects (control). Expression levels of EP1, EP3, DP1, and DP2 do not differ between EoE and controls. Values are mean ± SEM; Student’s t test; n = 6–7. * p

    Techniques Used: Expressing, Fluorescence

    ECIS ® cell impedance assays were performed in esophageal epithelial cells with 10 and 300 nM of EP4 agonist ONO-AE1-329 ( a ) and EP2 agonist butaprost ( b ) (or medium as control). The decrease in resistance correlates with a loss in the monolayer’s integrity indicative of cell barrier decrease. n = 5–9; values are mean ± SEM; one-way-ANOVA; Tukey’s post hoc test; * p
    Figure Legend Snippet: ECIS ® cell impedance assays were performed in esophageal epithelial cells with 10 and 300 nM of EP4 agonist ONO-AE1-329 ( a ) and EP2 agonist butaprost ( b ) (or medium as control). The decrease in resistance correlates with a loss in the monolayer’s integrity indicative of cell barrier decrease. n = 5–9; values are mean ± SEM; one-way-ANOVA; Tukey’s post hoc test; * p

    Techniques Used: Electric Cell-substrate Impedance Sensing

    2) Product Images from "Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide"

    Article Title: Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-016-0780-7

    Involvement of EP2 in the expression of genes related to classical microglial activation. mRNA expression of iNOS ( a , b ), NOX2 ( c , d ), G6PD ( e , f ), and VEGFA ( g , h ) in microglia. a , c , e , g LPS increases mRNA expression at 24 h, and an EP2 antagonist (1 μM PF 04418948) reduces the effect of LPS (three to four independent experiments). b , d , f , h An EP2 agonist (1 μM butaprost) increases the mRNA expression of these genes at 8 h (three independent experiments). * p
    Figure Legend Snippet: Involvement of EP2 in the expression of genes related to classical microglial activation. mRNA expression of iNOS ( a , b ), NOX2 ( c , d ), G6PD ( e , f ), and VEGFA ( g , h ) in microglia. a , c , e , g LPS increases mRNA expression at 24 h, and an EP2 antagonist (1 μM PF 04418948) reduces the effect of LPS (three to four independent experiments). b , d , f , h An EP2 agonist (1 μM butaprost) increases the mRNA expression of these genes at 8 h (three independent experiments). * p

    Techniques Used: Expressing, Activation Assay

    Effects of EP2 antagonist on LPS-induced protein expression in microglia. a LPS increases the expression of NOX2 protein at 24 h. b This effect is reduced by the EP2 antagonist (1 μM PF 04418948). c Also, LPS increases VEGFA protein expression and the EP2 antagonist attenuates this effect ( d ). e The EP2 antagonist reduces the nitrite production in the microglial culture medium induced by LPS at 48 h, indicating that EP2 contributes to LPS-induced iNOS activity. The figure illustrates a representative experiment ( n = 7–11) out of the three independent experiments. a , c n = 3–6 in two to four independent experiments; b , d n = 3 obtained in three independent experiments. * p ≤ 0.05; ** p
    Figure Legend Snippet: Effects of EP2 antagonist on LPS-induced protein expression in microglia. a LPS increases the expression of NOX2 protein at 24 h. b This effect is reduced by the EP2 antagonist (1 μM PF 04418948). c Also, LPS increases VEGFA protein expression and the EP2 antagonist attenuates this effect ( d ). e The EP2 antagonist reduces the nitrite production in the microglial culture medium induced by LPS at 48 h, indicating that EP2 contributes to LPS-induced iNOS activity. The figure illustrates a representative experiment ( n = 7–11) out of the three independent experiments. a , c n = 3–6 in two to four independent experiments; b , d n = 3 obtained in three independent experiments. * p ≤ 0.05; ** p

    Techniques Used: Expressing, Activity Assay

    Schematic representation of the suggested dynamic effects of PGE 2 on naïve and primed microglia. a Stimulation of naïve microglia with LPS activates TLR-4 leading to very rapid transcription of pro-inflammatory genes, including COX-2. COX-2 produces PGE 2 and increases the concentration of PGE 2 in the extracellular space. Although selective agonists of EP4 or EP2 exert anti-inflammatory effects, PGE 2 mainly induces anti-inflammatory effects through EP4 in naïve microglia. b During the course of classical microglial activation, EP4 receptor expression is down-regulated whereas EP2 expression increases and is found in peri-nuclear/nuclear zones enabling responses to high intracellular PGE 2 . EP2 favors the induction of iNOS, NOX2, and G6PD. G6PD is involved in metabolic changes promoting glucose utilization through the pentose pathway that generates NADPH to fuel NO production by iNOS and free radical generation by NOX2. AA arachidonic acid, COX2 cyclooxygenase-2, G6PD glucose-6-phosphate dehydrogenase, HK hexokinase, TLR-4 Toll-like receptor-4, TF transcription factors, TNF-α tumor necrosis factor-α
    Figure Legend Snippet: Schematic representation of the suggested dynamic effects of PGE 2 on naïve and primed microglia. a Stimulation of naïve microglia with LPS activates TLR-4 leading to very rapid transcription of pro-inflammatory genes, including COX-2. COX-2 produces PGE 2 and increases the concentration of PGE 2 in the extracellular space. Although selective agonists of EP4 or EP2 exert anti-inflammatory effects, PGE 2 mainly induces anti-inflammatory effects through EP4 in naïve microglia. b During the course of classical microglial activation, EP4 receptor expression is down-regulated whereas EP2 expression increases and is found in peri-nuclear/nuclear zones enabling responses to high intracellular PGE 2 . EP2 favors the induction of iNOS, NOX2, and G6PD. G6PD is involved in metabolic changes promoting glucose utilization through the pentose pathway that generates NADPH to fuel NO production by iNOS and free radical generation by NOX2. AA arachidonic acid, COX2 cyclooxygenase-2, G6PD glucose-6-phosphate dehydrogenase, HK hexokinase, TLR-4 Toll-like receptor-4, TF transcription factors, TNF-α tumor necrosis factor-α

    Techniques Used: Concentration Assay, Activation Assay, Expressing

    Effects of LPS on EP4 and EP2 mRNA expression. a , b LPS strongly depresses EP4 mRNA expression versus naïve cells, whereas it increases EP2 mRNA expression ( n = 6–7 in three independent cultures for each cell type) in microglia ( a ) and astroglia ( b ) at 8 h. The level of mRNA for each EP and cell type is expressed as fold versus the corresponding basal mRNA levels in non-stimulated cells (control). c The intensity of microglia EP4 immunoreactivity is strongly reduced 24 h after LPS exposure ( n = 4 in two independent experiments). d Microglia EP2 immunoreactivity is detected in a peri-nuclear/nuclear localization 24 h after LPS ( n = 3 replicates per treatment group in three independent experiments). e Western blotting shows increased EP2 expression after LPS in microglia and macrophages. f Quantification of EP2 protein expression in control microglia and 8 and 24 h after LPS, where values (EP2/GAPDH ratio) are expressed as fold versus control (* p
    Figure Legend Snippet: Effects of LPS on EP4 and EP2 mRNA expression. a , b LPS strongly depresses EP4 mRNA expression versus naïve cells, whereas it increases EP2 mRNA expression ( n = 6–7 in three independent cultures for each cell type) in microglia ( a ) and astroglia ( b ) at 8 h. The level of mRNA for each EP and cell type is expressed as fold versus the corresponding basal mRNA levels in non-stimulated cells (control). c The intensity of microglia EP4 immunoreactivity is strongly reduced 24 h after LPS exposure ( n = 4 in two independent experiments). d Microglia EP2 immunoreactivity is detected in a peri-nuclear/nuclear localization 24 h after LPS ( n = 3 replicates per treatment group in three independent experiments). e Western blotting shows increased EP2 expression after LPS in microglia and macrophages. f Quantification of EP2 protein expression in control microglia and 8 and 24 h after LPS, where values (EP2/GAPDH ratio) are expressed as fold versus control (* p

    Techniques Used: Expressing, Western Blot

    Relative EP mRNA expression in glial cells. a EP mRNA expression in naïve astroglia and microglia cultures ( n = 6–7 in three independent cultures for each cell type). For comparative purposes, the level of mRNA for each EP and cell type is expressed as fold versus mean EP1 mRNA levels in astroglia. EP4 and to a lower extent EP2 expression is comparatively higher than EP1 and EP3 in naïve microglia and is higher in microglia than astroglia (** p
    Figure Legend Snippet: Relative EP mRNA expression in glial cells. a EP mRNA expression in naïve astroglia and microglia cultures ( n = 6–7 in three independent cultures for each cell type). For comparative purposes, the level of mRNA for each EP and cell type is expressed as fold versus mean EP1 mRNA levels in astroglia. EP4 and to a lower extent EP2 expression is comparatively higher than EP1 and EP3 in naïve microglia and is higher in microglia than astroglia (** p

    Techniques Used: Expressing

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    Alomone Labs rrid ab 465844 rabbit polyclonal anti mouse ep2 fitc alomone
    Rrid Ab 465844 Rabbit Polyclonal Anti Mouse Ep2 Fitc Alomone, supplied by Alomone Labs, 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/rrid ab 465844 rabbit polyclonal anti mouse ep2 fitc alomone/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rrid ab 465844 rabbit polyclonal anti mouse ep2 fitc alomone - by Bioz Stars, 2022-11
    93/100 stars
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    Alomone Labs rabbit polyclonal antibodies against ep2
    Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of <t>EP2</t> agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values
    Rabbit Polyclonal Antibodies Against Ep2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal antibodies against ep2/product/Alomone Labs
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal antibodies against ep2 - by Bioz Stars, 2022-11
    91/100 stars
      Buy from Supplier

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    Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of EP2 agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values

    Journal: Digestive Diseases and Sciences

    Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

    doi: 10.1007/s10620-019-05623-5

    Figure Lengend Snippet: Chemotaxis and adhesion assays with human blood eosinophils. Eosinophils from healthy donors were incubated with 100 nM of EP4 agonist ONO-AE1-329 or with 100 nM of the EP4 antagonist ONO-AE3-208 ( a ), with 100 and 300 nM of EP2 agonist butaprost ( b ), or with 100 nM of DP1 agonist BW245C and 1 µM of DP1 antagonist MK0524 ( c ), and chemotaxis was assessed using esophageal epithelial cell supernatant for chemoattraction. n = 6–12 (for a , and b ) and n = 3 ( c ); values are mean ± SEM; one-way ANOVA; Tukey’s post hoc test. *** p values

    Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

    Techniques: Chemotaxis Assay, Incubation

    Immunohistochemistry shows strong DP1 staining in eosinophils ( a , arrows ), while epithelial cells in esophageal mucosal biopsies are practically devoid of staining. EP4 ( b ) and EP2 ( c ) immunostaining is prominent in eosinophils ( arrows ) but also visible in epithelial cells ( arrowheads ). 3-3′-Diaminobenzidine (DAB) was used as visualization substrate for DP1 and EP4 staining (brown precipitates), while for EP2, ImmPact™ NovaRED substrate was used (red precipitate). Calibration bar: 50 µm

    Journal: Digestive Diseases and Sciences

    Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

    doi: 10.1007/s10620-019-05623-5

    Figure Lengend Snippet: Immunohistochemistry shows strong DP1 staining in eosinophils ( a , arrows ), while epithelial cells in esophageal mucosal biopsies are practically devoid of staining. EP4 ( b ) and EP2 ( c ) immunostaining is prominent in eosinophils ( arrows ) but also visible in epithelial cells ( arrowheads ). 3-3′-Diaminobenzidine (DAB) was used as visualization substrate for DP1 and EP4 staining (brown precipitates), while for EP2, ImmPact™ NovaRED substrate was used (red precipitate). Calibration bar: 50 µm

    Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

    Techniques: Immunohistochemistry, Staining, Immunostaining

    Flow cytometric evaluation of EP (EP1–4) and DP (DP1, DP2) receptor expression in human blood eosinophils. Changes in receptor expression were recorded as mean fluorescence intensity and data are expressed as fold increase in fluorescence over isotype control. Expression levels of EP2 and EP4 are significantly lower in patients with eosinophilic esophagitis (EoE) compared to healthy control subjects (control). Expression levels of EP1, EP3, DP1, and DP2 do not differ between EoE and controls. Values are mean ± SEM; Student’s t test; n = 6–7. * p

    Journal: Digestive Diseases and Sciences

    Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

    doi: 10.1007/s10620-019-05623-5

    Figure Lengend Snippet: Flow cytometric evaluation of EP (EP1–4) and DP (DP1, DP2) receptor expression in human blood eosinophils. Changes in receptor expression were recorded as mean fluorescence intensity and data are expressed as fold increase in fluorescence over isotype control. Expression levels of EP2 and EP4 are significantly lower in patients with eosinophilic esophagitis (EoE) compared to healthy control subjects (control). Expression levels of EP1, EP3, DP1, and DP2 do not differ between EoE and controls. Values are mean ± SEM; Student’s t test; n = 6–7. * p

    Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

    Techniques: Expressing, Fluorescence

    ECIS ® cell impedance assays were performed in esophageal epithelial cells with 10 and 300 nM of EP4 agonist ONO-AE1-329 ( a ) and EP2 agonist butaprost ( b ) (or medium as control). The decrease in resistance correlates with a loss in the monolayer’s integrity indicative of cell barrier decrease. n = 5–9; values are mean ± SEM; one-way-ANOVA; Tukey’s post hoc test; * p

    Journal: Digestive Diseases and Sciences

    Article Title: Involvement of EP2 and EP4 Receptors in Eosinophilic Esophagitis: A Pilot Study

    doi: 10.1007/s10620-019-05623-5

    Figure Lengend Snippet: ECIS ® cell impedance assays were performed in esophageal epithelial cells with 10 and 300 nM of EP4 agonist ONO-AE1-329 ( a ) and EP2 agonist butaprost ( b ) (or medium as control). The decrease in resistance correlates with a loss in the monolayer’s integrity indicative of cell barrier decrease. n = 5–9; values are mean ± SEM; one-way-ANOVA; Tukey’s post hoc test; * p

    Article Snippet: For flow cytometric visualization of the EP receptors, cells were incubated with rabbit polyclonal antibodies against EP1, EP2, and EP3 (20 μg/ml; Alomone Labs, Jerusalem, Israel) and mouse polyclonal antibody against EP4 (20 μg/ml; Alomone Labs, Jerusalem, Israel), or normal rabbit or mouse IgG (20 μg/ml; Santa Cruz Biotechnology, Dallas, TX, USA) for 30 min at 4 °C.

    Techniques: Electric Cell-substrate Impedance Sensing

    Involvement of EP2 in the expression of genes related to classical microglial activation. mRNA expression of iNOS ( a , b ), NOX2 ( c , d ), G6PD ( e , f ), and VEGFA ( g , h ) in microglia. a , c , e , g LPS increases mRNA expression at 24 h, and an EP2 antagonist (1 μM PF 04418948) reduces the effect of LPS (three to four independent experiments). b , d , f , h An EP2 agonist (1 μM butaprost) increases the mRNA expression of these genes at 8 h (three independent experiments). * p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide

    doi: 10.1186/s12974-016-0780-7

    Figure Lengend Snippet: Involvement of EP2 in the expression of genes related to classical microglial activation. mRNA expression of iNOS ( a , b ), NOX2 ( c , d ), G6PD ( e , f ), and VEGFA ( g , h ) in microglia. a , c , e , g LPS increases mRNA expression at 24 h, and an EP2 antagonist (1 μM PF 04418948) reduces the effect of LPS (three to four independent experiments). b , d , f , h An EP2 agonist (1 μM butaprost) increases the mRNA expression of these genes at 8 h (three independent experiments). * p

    Article Snippet: The cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min, blocked with 3% goat serum in PBS for 1 h and incubated overnight at 4 °C with the primary rabbit antibodies against the EP2 (#APR-064, kindly provided by Alomone Labs) (1:100) and EP4 (#ab93486, Abcam, Cambridge, UK) (1:200) receptors.

    Techniques: Expressing, Activation Assay

    Effects of EP2 antagonist on LPS-induced protein expression in microglia. a LPS increases the expression of NOX2 protein at 24 h. b This effect is reduced by the EP2 antagonist (1 μM PF 04418948). c Also, LPS increases VEGFA protein expression and the EP2 antagonist attenuates this effect ( d ). e The EP2 antagonist reduces the nitrite production in the microglial culture medium induced by LPS at 48 h, indicating that EP2 contributes to LPS-induced iNOS activity. The figure illustrates a representative experiment ( n = 7–11) out of the three independent experiments. a , c n = 3–6 in two to four independent experiments; b , d n = 3 obtained in three independent experiments. * p ≤ 0.05; ** p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide

    doi: 10.1186/s12974-016-0780-7

    Figure Lengend Snippet: Effects of EP2 antagonist on LPS-induced protein expression in microglia. a LPS increases the expression of NOX2 protein at 24 h. b This effect is reduced by the EP2 antagonist (1 μM PF 04418948). c Also, LPS increases VEGFA protein expression and the EP2 antagonist attenuates this effect ( d ). e The EP2 antagonist reduces the nitrite production in the microglial culture medium induced by LPS at 48 h, indicating that EP2 contributes to LPS-induced iNOS activity. The figure illustrates a representative experiment ( n = 7–11) out of the three independent experiments. a , c n = 3–6 in two to four independent experiments; b , d n = 3 obtained in three independent experiments. * p ≤ 0.05; ** p

    Article Snippet: The cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min, blocked with 3% goat serum in PBS for 1 h and incubated overnight at 4 °C with the primary rabbit antibodies against the EP2 (#APR-064, kindly provided by Alomone Labs) (1:100) and EP4 (#ab93486, Abcam, Cambridge, UK) (1:200) receptors.

    Techniques: Expressing, Activity Assay

    Schematic representation of the suggested dynamic effects of PGE 2 on naïve and primed microglia. a Stimulation of naïve microglia with LPS activates TLR-4 leading to very rapid transcription of pro-inflammatory genes, including COX-2. COX-2 produces PGE 2 and increases the concentration of PGE 2 in the extracellular space. Although selective agonists of EP4 or EP2 exert anti-inflammatory effects, PGE 2 mainly induces anti-inflammatory effects through EP4 in naïve microglia. b During the course of classical microglial activation, EP4 receptor expression is down-regulated whereas EP2 expression increases and is found in peri-nuclear/nuclear zones enabling responses to high intracellular PGE 2 . EP2 favors the induction of iNOS, NOX2, and G6PD. G6PD is involved in metabolic changes promoting glucose utilization through the pentose pathway that generates NADPH to fuel NO production by iNOS and free radical generation by NOX2. AA arachidonic acid, COX2 cyclooxygenase-2, G6PD glucose-6-phosphate dehydrogenase, HK hexokinase, TLR-4 Toll-like receptor-4, TF transcription factors, TNF-α tumor necrosis factor-α

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide

    doi: 10.1186/s12974-016-0780-7

    Figure Lengend Snippet: Schematic representation of the suggested dynamic effects of PGE 2 on naïve and primed microglia. a Stimulation of naïve microglia with LPS activates TLR-4 leading to very rapid transcription of pro-inflammatory genes, including COX-2. COX-2 produces PGE 2 and increases the concentration of PGE 2 in the extracellular space. Although selective agonists of EP4 or EP2 exert anti-inflammatory effects, PGE 2 mainly induces anti-inflammatory effects through EP4 in naïve microglia. b During the course of classical microglial activation, EP4 receptor expression is down-regulated whereas EP2 expression increases and is found in peri-nuclear/nuclear zones enabling responses to high intracellular PGE 2 . EP2 favors the induction of iNOS, NOX2, and G6PD. G6PD is involved in metabolic changes promoting glucose utilization through the pentose pathway that generates NADPH to fuel NO production by iNOS and free radical generation by NOX2. AA arachidonic acid, COX2 cyclooxygenase-2, G6PD glucose-6-phosphate dehydrogenase, HK hexokinase, TLR-4 Toll-like receptor-4, TF transcription factors, TNF-α tumor necrosis factor-α

    Article Snippet: The cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min, blocked with 3% goat serum in PBS for 1 h and incubated overnight at 4 °C with the primary rabbit antibodies against the EP2 (#APR-064, kindly provided by Alomone Labs) (1:100) and EP4 (#ab93486, Abcam, Cambridge, UK) (1:200) receptors.

    Techniques: Concentration Assay, Activation Assay, Expressing

    Effects of LPS on EP4 and EP2 mRNA expression. a , b LPS strongly depresses EP4 mRNA expression versus naïve cells, whereas it increases EP2 mRNA expression ( n = 6–7 in three independent cultures for each cell type) in microglia ( a ) and astroglia ( b ) at 8 h. The level of mRNA for each EP and cell type is expressed as fold versus the corresponding basal mRNA levels in non-stimulated cells (control). c The intensity of microglia EP4 immunoreactivity is strongly reduced 24 h after LPS exposure ( n = 4 in two independent experiments). d Microglia EP2 immunoreactivity is detected in a peri-nuclear/nuclear localization 24 h after LPS ( n = 3 replicates per treatment group in three independent experiments). e Western blotting shows increased EP2 expression after LPS in microglia and macrophages. f Quantification of EP2 protein expression in control microglia and 8 and 24 h after LPS, where values (EP2/GAPDH ratio) are expressed as fold versus control (* p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide

    doi: 10.1186/s12974-016-0780-7

    Figure Lengend Snippet: Effects of LPS on EP4 and EP2 mRNA expression. a , b LPS strongly depresses EP4 mRNA expression versus naïve cells, whereas it increases EP2 mRNA expression ( n = 6–7 in three independent cultures for each cell type) in microglia ( a ) and astroglia ( b ) at 8 h. The level of mRNA for each EP and cell type is expressed as fold versus the corresponding basal mRNA levels in non-stimulated cells (control). c The intensity of microglia EP4 immunoreactivity is strongly reduced 24 h after LPS exposure ( n = 4 in two independent experiments). d Microglia EP2 immunoreactivity is detected in a peri-nuclear/nuclear localization 24 h after LPS ( n = 3 replicates per treatment group in three independent experiments). e Western blotting shows increased EP2 expression after LPS in microglia and macrophages. f Quantification of EP2 protein expression in control microglia and 8 and 24 h after LPS, where values (EP2/GAPDH ratio) are expressed as fold versus control (* p

    Article Snippet: The cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min, blocked with 3% goat serum in PBS for 1 h and incubated overnight at 4 °C with the primary rabbit antibodies against the EP2 (#APR-064, kindly provided by Alomone Labs) (1:100) and EP4 (#ab93486, Abcam, Cambridge, UK) (1:200) receptors.

    Techniques: Expressing, Western Blot

    Relative EP mRNA expression in glial cells. a EP mRNA expression in naïve astroglia and microglia cultures ( n = 6–7 in three independent cultures for each cell type). For comparative purposes, the level of mRNA for each EP and cell type is expressed as fold versus mean EP1 mRNA levels in astroglia. EP4 and to a lower extent EP2 expression is comparatively higher than EP1 and EP3 in naïve microglia and is higher in microglia than astroglia (** p

    Journal: Journal of Neuroinflammation

    Article Title: Differential expression of E-type prostanoid receptors 2 and 4 in microglia stimulated with lipopolysaccharide

    doi: 10.1186/s12974-016-0780-7

    Figure Lengend Snippet: Relative EP mRNA expression in glial cells. a EP mRNA expression in naïve astroglia and microglia cultures ( n = 6–7 in three independent cultures for each cell type). For comparative purposes, the level of mRNA for each EP and cell type is expressed as fold versus mean EP1 mRNA levels in astroglia. EP4 and to a lower extent EP2 expression is comparatively higher than EP1 and EP3 in naïve microglia and is higher in microglia than astroglia (** p

    Article Snippet: The cells were fixed in 4% paraformaldehyde for 20 min, permeabilized with 0.2% Triton X-100 (Sigma) in PBS for 10 min, blocked with 3% goat serum in PBS for 1 h and incubated overnight at 4 °C with the primary rabbit antibodies against the EP2 (#APR-064, kindly provided by Alomone Labs) (1:100) and EP4 (#ab93486, Abcam, Cambridge, UK) (1:200) receptors.

    Techniques: Expressing