anti h1r  (Alomone Labs)


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    Alomone Labs anti h1r
    Effects of HR antagonists on HA-induced suppression of astrocytic TNF-α and IL-1β production. a Incubation with histamine at the doses of 0.001, 0.01, 0.1, and 1 μg/ml for 24 h produced a concentration-dependent suppression of TNF-α and IL-1β secretion from primary cultured astrocytes. The <t>H1R</t> antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) were added to astrocytes alone or 30 min before addition of histamine (0.1 μg/ml) for 24 h. b Time courses of suppression of TNF-α and IL-1β release by histamine. Histamine at 0.1 μg/ml was incubated with astrocytes at 37 °C for 2, 6, 12, 24, 48, and 72 h. c The levels of TNF-α and IL-1β mRNA expression were analyzed by quantitative RT-PCR. * p
    Anti H1r, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti h1r/product/Alomone Labs
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    anti h1r - by Bioz Stars, 2022-12
    91/100 stars

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    1) Product Images from "Histamine upregulates the expression of histamine receptors and increases the neuroprotective effect of astrocytes"

    Article Title: Histamine upregulates the expression of histamine receptors and increases the neuroprotective effect of astrocytes

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-018-1068-x

    Effects of HR antagonists on HA-induced suppression of astrocytic TNF-α and IL-1β production. a Incubation with histamine at the doses of 0.001, 0.01, 0.1, and 1 μg/ml for 24 h produced a concentration-dependent suppression of TNF-α and IL-1β secretion from primary cultured astrocytes. The H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) were added to astrocytes alone or 30 min before addition of histamine (0.1 μg/ml) for 24 h. b Time courses of suppression of TNF-α and IL-1β release by histamine. Histamine at 0.1 μg/ml was incubated with astrocytes at 37 °C for 2, 6, 12, 24, 48, and 72 h. c The levels of TNF-α and IL-1β mRNA expression were analyzed by quantitative RT-PCR. * p
    Figure Legend Snippet: Effects of HR antagonists on HA-induced suppression of astrocytic TNF-α and IL-1β production. a Incubation with histamine at the doses of 0.001, 0.01, 0.1, and 1 μg/ml for 24 h produced a concentration-dependent suppression of TNF-α and IL-1β secretion from primary cultured astrocytes. The H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) were added to astrocytes alone or 30 min before addition of histamine (0.1 μg/ml) for 24 h. b Time courses of suppression of TNF-α and IL-1β release by histamine. Histamine at 0.1 μg/ml was incubated with astrocytes at 37 °C for 2, 6, 12, 24, 48, and 72 h. c The levels of TNF-α and IL-1β mRNA expression were analyzed by quantitative RT-PCR. * p

    Techniques Used: Incubation, Produced, Concentration Assay, Cell Culture, Expressing, Quantitative RT-PCR

    Upregulation of H1R, H2R, and H3R expression in primary cultured astrocytes by histamine. Primary astrocytes were stimulated with histamine at 0.1 μg/ml for 24 h. a – c Immunofluorescence analysis of GFAP and histamine receptor expression. The cells were stained for the astrocyte marker GFAP (red) and for H1R, H2R, and H3R (green). Upregulated GFAP and HR expression on activated astrocytes was observed using confocal microscopy. The blue staining represents DAPI. Scale bar = 25 μm. d The expression levels of the histamine H1, H2, and H3 receptor subtypes were detected via Western blotting using specific antibodies. Each blot is representative of three experiments. e The expression levels of the histamine H1, H2, and H3 receptor subtypes were examined by quantitative RT-PCR. * p
    Figure Legend Snippet: Upregulation of H1R, H2R, and H3R expression in primary cultured astrocytes by histamine. Primary astrocytes were stimulated with histamine at 0.1 μg/ml for 24 h. a – c Immunofluorescence analysis of GFAP and histamine receptor expression. The cells were stained for the astrocyte marker GFAP (red) and for H1R, H2R, and H3R (green). Upregulated GFAP and HR expression on activated astrocytes was observed using confocal microscopy. The blue staining represents DAPI. Scale bar = 25 μm. d The expression levels of the histamine H1, H2, and H3 receptor subtypes were detected via Western blotting using specific antibodies. Each blot is representative of three experiments. e The expression levels of the histamine H1, H2, and H3 receptor subtypes were examined by quantitative RT-PCR. * p

    Techniques Used: Expressing, Cell Culture, Immunofluorescence, Staining, Marker, Confocal Microscopy, Western Blot, Quantitative RT-PCR

    Identification of expression of histamine receptors in primary cultured astrocytes. a Immunolocalization of H1R, H2R, H3R, and H4R in astrocytes was performed by using antibodies against H1R, H2R, H3R, and H4R (green) and an antibody against the astrocyte marker GFAP (red). The results were imaged with a laser scanning confocal microscope. The blue staining represents DAPI. Scale bar = 25 μm. b Western blotting analysis of H1R, H2R, H3R, and H4R in the extracts of rat astrocytes. c The expression levels of the histamine H1, H2, H3, and H4 receptor subtypes were examined by quantitative RT-PCR. The data are presented as the mean ± s.e.m. of three independent experiments
    Figure Legend Snippet: Identification of expression of histamine receptors in primary cultured astrocytes. a Immunolocalization of H1R, H2R, H3R, and H4R in astrocytes was performed by using antibodies against H1R, H2R, H3R, and H4R (green) and an antibody against the astrocyte marker GFAP (red). The results were imaged with a laser scanning confocal microscope. The blue staining represents DAPI. Scale bar = 25 μm. b Western blotting analysis of H1R, H2R, H3R, and H4R in the extracts of rat astrocytes. c The expression levels of the histamine H1, H2, H3, and H4 receptor subtypes were examined by quantitative RT-PCR. The data are presented as the mean ± s.e.m. of three independent experiments

    Techniques Used: Expressing, Cell Culture, Marker, Microscopy, Staining, Western Blot, Quantitative RT-PCR

    Effects of HR antagonists on HA-induced stimulation of GDNF protein levels in astrocytes. a Incubation with histamine at the doses of 0.01 and 0.1 μg/ml for 6 and 24 h significantly promoted GDNF expression in astrocytes. b Astrocyte cells were stimulated with histamine (0.1 μg/ml) in the absence or presence of the H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) added 30 min before stimulation. At 24 h after the addition of histamine, levels of GDNF were detected by Western blotting, quantified, and normalized to GAPDH levels. Values are expressed relative to the control, which was set to 1. ** p
    Figure Legend Snippet: Effects of HR antagonists on HA-induced stimulation of GDNF protein levels in astrocytes. a Incubation with histamine at the doses of 0.01 and 0.1 μg/ml for 6 and 24 h significantly promoted GDNF expression in astrocytes. b Astrocyte cells were stimulated with histamine (0.1 μg/ml) in the absence or presence of the H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) added 30 min before stimulation. At 24 h after the addition of histamine, levels of GDNF were detected by Western blotting, quantified, and normalized to GAPDH levels. Values are expressed relative to the control, which was set to 1. ** p

    Techniques Used: Incubation, Expressing, Western Blot

    2) Product Images from "Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis"

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.01689

    Histamine modulates inflammation in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting (WB) and immunoreactions with anti-NADPH oxidase 2 (NOX2) (A) , anti-ARG-1 (B) . SOD1-G93A microglia stimulated with histamine (100 µM) in the presence of H1R–H2R–H3R–H4R antagonists for 6 h were subjected to immunoreactions with anti-NOX2 (C) or anti-ARG-1 (D) . SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to WB and immunoreactions with anti-CD163 (E) or anti-CD206 (F) . Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p
    Figure Legend Snippet: Histamine modulates inflammation in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting (WB) and immunoreactions with anti-NADPH oxidase 2 (NOX2) (A) , anti-ARG-1 (B) . SOD1-G93A microglia stimulated with histamine (100 µM) in the presence of H1R–H2R–H3R–H4R antagonists for 6 h were subjected to immunoreactions with anti-NOX2 (C) or anti-ARG-1 (D) . SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to WB and immunoreactions with anti-CD163 (E) or anti-CD206 (F) . Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Techniques Used: Western Blot

    Protein expression levels of H1-H4R in cortex and lumbar spinal cord from wild-type (WT) and SOD1-G93A mice. Cortex (A,C,E,G) and lumbar spinal cord (B,D,F,H) from WT (~20 weeks) and SOD1-G93A mice at different stages of disease (100, 140 days, and end stage) ( n = 4/group) were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to WT mice, * p
    Figure Legend Snippet: Protein expression levels of H1-H4R in cortex and lumbar spinal cord from wild-type (WT) and SOD1-G93A mice. Cortex (A,C,E,G) and lumbar spinal cord (B,D,F,H) from WT (~20 weeks) and SOD1-G93A mice at different stages of disease (100, 140 days, and end stage) ( n = 4/group) were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to WT mice, * p

    Techniques Used: Expressing, Mouse Assay, SDS Page, Western Blot

    MAPKs are activated by histamine in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were treated with histamine (100 µM) for 15 min–1 h–6 h and equal amounts of total lysates were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-p-p38 and anti-p-38 (A) or with anti-pERK and anti-ERK (B) . (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 1 h in the presence of specific antagonists for H1R (orphenadrine, 10 µM), H2R (ranitidine, 10 µM), H3R (thioperamide, 5 µM), or H4R (JNJ-7777120, 5 µM) were subjected to immunoreactions with anti-pERK and anti-ERK. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p
    Figure Legend Snippet: MAPKs are activated by histamine in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were treated with histamine (100 µM) for 15 min–1 h–6 h and equal amounts of total lysates were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-p-p38 and anti-p-38 (A) or with anti-pERK and anti-ERK (B) . (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 1 h in the presence of specific antagonists for H1R (orphenadrine, 10 µM), H2R (ranitidine, 10 µM), H3R (thioperamide, 5 µM), or H4R (JNJ-7777120, 5 µM) were subjected to immunoreactions with anti-pERK and anti-ERK. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Techniques Used: SDS Page, Western Blot

    NF-κB pathway is modulated by histamine in SOD1-G93A microglia. (A) Total RNA was extracted from SOD1-G93A microglia stimulated with histamine (100 µM) for 6–18 h and the expression profiles of IL-10, IL-6, and IL-1β were examined by qRT-PCR. (B) Wild-type (WT) and SOD1-G93A microglia exposed to 100 µM histamine for 1–6–18 h were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 6 h in the presence of H1R–H2R–H3R–H4R antagonists were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (D) SOD1-G93A microglia treated with histamine (100 µM) for 6 h in the presence of specific inhibitors of pERK (PD98059, 100 µM) and pp38 (SB203580, 20 µM) were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p
    Figure Legend Snippet: NF-κB pathway is modulated by histamine in SOD1-G93A microglia. (A) Total RNA was extracted from SOD1-G93A microglia stimulated with histamine (100 µM) for 6–18 h and the expression profiles of IL-10, IL-6, and IL-1β were examined by qRT-PCR. (B) Wild-type (WT) and SOD1-G93A microglia exposed to 100 µM histamine for 1–6–18 h were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 6 h in the presence of H1R–H2R–H3R–H4R antagonists were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (D) SOD1-G93A microglia treated with histamine (100 µM) for 6 h in the presence of specific inhibitors of pERK (PD98059, 100 µM) and pp38 (SB203580, 20 µM) were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Techniques Used: Expressing, Quantitative RT-PCR

    Histamine induces migration in SOD1-G93A microglia. (A) Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting and immunoreactions with anti-P2Y12. Anti-GAPDH was used for protein normalization. (B) SOD1-G93A microglia were stimulated with 100 µM histamine in the presence or absence of H1R–H2R–H3R–H4R antagonists or P2Y12 antagonist (MRS2395, 10 µM) for 18 h and stained with anti-CD11b (green). Scale bar 50 µm. The number of migrating cells was then quantified. (C) WT microglia were stimulated with 100 microM histamine in the presence or absence of H1R-H2R-H3R-H4R antagonists for 18 h and the number of migrating cells was quantified, HA, histamine. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p
    Figure Legend Snippet: Histamine induces migration in SOD1-G93A microglia. (A) Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting and immunoreactions with anti-P2Y12. Anti-GAPDH was used for protein normalization. (B) SOD1-G93A microglia were stimulated with 100 µM histamine in the presence or absence of H1R–H2R–H3R–H4R antagonists or P2Y12 antagonist (MRS2395, 10 µM) for 18 h and stained with anti-CD11b (green). Scale bar 50 µm. The number of migrating cells was then quantified. (C) WT microglia were stimulated with 100 microM histamine in the presence or absence of H1R-H2R-H3R-H4R antagonists for 18 h and the number of migrating cells was quantified, HA, histamine. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Techniques Used: Migration, Western Blot, Staining

    SOD1-G93A microglia express histaminergic receptors. (A) Primary microglia from SOD1-G93A mice were stained with anti-CD11b (red) and anti-H1R, anti-H2R, anti-H3R, and anti-H4R (green). Höechst 33258 was used for nuclei (scale bar 50 µm). In inserts, wild-type (WT) microglia (scale bar 50 µm). (B) Equal amounts of WT and SOD1-G93A primary microglia total lysates were subjected to western blotting with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by Student’s t -test, as referred to WT cells, * p
    Figure Legend Snippet: SOD1-G93A microglia express histaminergic receptors. (A) Primary microglia from SOD1-G93A mice were stained with anti-CD11b (red) and anti-H1R, anti-H2R, anti-H3R, and anti-H4R (green). Höechst 33258 was used for nuclei (scale bar 50 µm). In inserts, wild-type (WT) microglia (scale bar 50 µm). (B) Equal amounts of WT and SOD1-G93A primary microglia total lysates were subjected to western blotting with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by Student’s t -test, as referred to WT cells, * p

    Techniques Used: Mouse Assay, Staining, Western Blot

    3) Product Images from "Chronic Ingestion of H1-Antihistamines Increase Progression of Atherosclerosis in Apolipoprotein E-/- Mice"

    Article Title: Chronic Ingestion of H1-Antihistamines Increase Progression of Atherosclerosis in Apolipoprotein E-/- Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0102165

    Effect of H1-antihistamines on H1R expression in atherosclerotic plaques. Atherosclerotic lesions around the aortic root of ApoE −/− mice were subjected to immunohistochemical analyses for H1R expression with anti-H1R (green) and DAPI (nuclear stain-blue). Panel (A) represents immunoflourescent staining for placebo (Pl), cetirizine low (C.L), cetirizine high (C.H), fexofenadine low (F.L) and fexofenadine high (F.H). Panel (B) depicts the integrated density of H1R staining. Magnification 40× at objective. n = 4.
    Figure Legend Snippet: Effect of H1-antihistamines on H1R expression in atherosclerotic plaques. Atherosclerotic lesions around the aortic root of ApoE −/− mice were subjected to immunohistochemical analyses for H1R expression with anti-H1R (green) and DAPI (nuclear stain-blue). Panel (A) represents immunoflourescent staining for placebo (Pl), cetirizine low (C.L), cetirizine high (C.H), fexofenadine low (F.L) and fexofenadine high (F.H). Panel (B) depicts the integrated density of H1R staining. Magnification 40× at objective. n = 4.

    Techniques Used: Expressing, Mouse Assay, Immunohistochemistry, Staining

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    Alomone Labs anti h1r
    Effects of HR antagonists on HA-induced suppression of astrocytic TNF-α and IL-1β production. a Incubation with histamine at the doses of 0.001, 0.01, 0.1, and 1 μg/ml for 24 h produced a concentration-dependent suppression of TNF-α and IL-1β secretion from primary cultured astrocytes. The <t>H1R</t> antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) were added to astrocytes alone or 30 min before addition of histamine (0.1 μg/ml) for 24 h. b Time courses of suppression of TNF-α and IL-1β release by histamine. Histamine at 0.1 μg/ml was incubated with astrocytes at 37 °C for 2, 6, 12, 24, 48, and 72 h. c The levels of TNF-α and IL-1β mRNA expression were analyzed by quantitative RT-PCR. * p
    Anti H1r, 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/anti h1r/product/Alomone Labs
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti h1r - by Bioz Stars, 2022-12
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    Effects of HR antagonists on HA-induced suppression of astrocytic TNF-α and IL-1β production. a Incubation with histamine at the doses of 0.001, 0.01, 0.1, and 1 μg/ml for 24 h produced a concentration-dependent suppression of TNF-α and IL-1β secretion from primary cultured astrocytes. The H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) were added to astrocytes alone or 30 min before addition of histamine (0.1 μg/ml) for 24 h. b Time courses of suppression of TNF-α and IL-1β release by histamine. Histamine at 0.1 μg/ml was incubated with astrocytes at 37 °C for 2, 6, 12, 24, 48, and 72 h. c The levels of TNF-α and IL-1β mRNA expression were analyzed by quantitative RT-PCR. * p

    Journal: Journal of Neuroinflammation

    Article Title: Histamine upregulates the expression of histamine receptors and increases the neuroprotective effect of astrocytes

    doi: 10.1186/s12974-018-1068-x

    Figure Lengend Snippet: Effects of HR antagonists on HA-induced suppression of astrocytic TNF-α and IL-1β production. a Incubation with histamine at the doses of 0.001, 0.01, 0.1, and 1 μg/ml for 24 h produced a concentration-dependent suppression of TNF-α and IL-1β secretion from primary cultured astrocytes. The H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) were added to astrocytes alone or 30 min before addition of histamine (0.1 μg/ml) for 24 h. b Time courses of suppression of TNF-α and IL-1β release by histamine. Histamine at 0.1 μg/ml was incubated with astrocytes at 37 °C for 2, 6, 12, 24, 48, and 72 h. c The levels of TNF-α and IL-1β mRNA expression were analyzed by quantitative RT-PCR. * p

    Article Snippet: Specific rabbit polyclonal anti-H1 receptor and rabbit polyclonal anti-H2 receptor antibodies were purchased from Alomone Labs Ltd. (Israel), and rabbit polyclonal anti-H4 receptor antibody was purchased from Santa Cruz (Santa Cruz Biotechnology, USA).

    Techniques: Incubation, Produced, Concentration Assay, Cell Culture, Expressing, Quantitative RT-PCR

    Upregulation of H1R, H2R, and H3R expression in primary cultured astrocytes by histamine. Primary astrocytes were stimulated with histamine at 0.1 μg/ml for 24 h. a – c Immunofluorescence analysis of GFAP and histamine receptor expression. The cells were stained for the astrocyte marker GFAP (red) and for H1R, H2R, and H3R (green). Upregulated GFAP and HR expression on activated astrocytes was observed using confocal microscopy. The blue staining represents DAPI. Scale bar = 25 μm. d The expression levels of the histamine H1, H2, and H3 receptor subtypes were detected via Western blotting using specific antibodies. Each blot is representative of three experiments. e The expression levels of the histamine H1, H2, and H3 receptor subtypes were examined by quantitative RT-PCR. * p

    Journal: Journal of Neuroinflammation

    Article Title: Histamine upregulates the expression of histamine receptors and increases the neuroprotective effect of astrocytes

    doi: 10.1186/s12974-018-1068-x

    Figure Lengend Snippet: Upregulation of H1R, H2R, and H3R expression in primary cultured astrocytes by histamine. Primary astrocytes were stimulated with histamine at 0.1 μg/ml for 24 h. a – c Immunofluorescence analysis of GFAP and histamine receptor expression. The cells were stained for the astrocyte marker GFAP (red) and for H1R, H2R, and H3R (green). Upregulated GFAP and HR expression on activated astrocytes was observed using confocal microscopy. The blue staining represents DAPI. Scale bar = 25 μm. d The expression levels of the histamine H1, H2, and H3 receptor subtypes were detected via Western blotting using specific antibodies. Each blot is representative of three experiments. e The expression levels of the histamine H1, H2, and H3 receptor subtypes were examined by quantitative RT-PCR. * p

    Article Snippet: Specific rabbit polyclonal anti-H1 receptor and rabbit polyclonal anti-H2 receptor antibodies were purchased from Alomone Labs Ltd. (Israel), and rabbit polyclonal anti-H4 receptor antibody was purchased from Santa Cruz (Santa Cruz Biotechnology, USA).

    Techniques: Expressing, Cell Culture, Immunofluorescence, Staining, Marker, Confocal Microscopy, Western Blot, Quantitative RT-PCR

    Identification of expression of histamine receptors in primary cultured astrocytes. a Immunolocalization of H1R, H2R, H3R, and H4R in astrocytes was performed by using antibodies against H1R, H2R, H3R, and H4R (green) and an antibody against the astrocyte marker GFAP (red). The results were imaged with a laser scanning confocal microscope. The blue staining represents DAPI. Scale bar = 25 μm. b Western blotting analysis of H1R, H2R, H3R, and H4R in the extracts of rat astrocytes. c The expression levels of the histamine H1, H2, H3, and H4 receptor subtypes were examined by quantitative RT-PCR. The data are presented as the mean ± s.e.m. of three independent experiments

    Journal: Journal of Neuroinflammation

    Article Title: Histamine upregulates the expression of histamine receptors and increases the neuroprotective effect of astrocytes

    doi: 10.1186/s12974-018-1068-x

    Figure Lengend Snippet: Identification of expression of histamine receptors in primary cultured astrocytes. a Immunolocalization of H1R, H2R, H3R, and H4R in astrocytes was performed by using antibodies against H1R, H2R, H3R, and H4R (green) and an antibody against the astrocyte marker GFAP (red). The results were imaged with a laser scanning confocal microscope. The blue staining represents DAPI. Scale bar = 25 μm. b Western blotting analysis of H1R, H2R, H3R, and H4R in the extracts of rat astrocytes. c The expression levels of the histamine H1, H2, H3, and H4 receptor subtypes were examined by quantitative RT-PCR. The data are presented as the mean ± s.e.m. of three independent experiments

    Article Snippet: Specific rabbit polyclonal anti-H1 receptor and rabbit polyclonal anti-H2 receptor antibodies were purchased from Alomone Labs Ltd. (Israel), and rabbit polyclonal anti-H4 receptor antibody was purchased from Santa Cruz (Santa Cruz Biotechnology, USA).

    Techniques: Expressing, Cell Culture, Marker, Microscopy, Staining, Western Blot, Quantitative RT-PCR

    Effects of HR antagonists on HA-induced stimulation of GDNF protein levels in astrocytes. a Incubation with histamine at the doses of 0.01 and 0.1 μg/ml for 6 and 24 h significantly promoted GDNF expression in astrocytes. b Astrocyte cells were stimulated with histamine (0.1 μg/ml) in the absence or presence of the H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) added 30 min before stimulation. At 24 h after the addition of histamine, levels of GDNF were detected by Western blotting, quantified, and normalized to GAPDH levels. Values are expressed relative to the control, which was set to 1. ** p

    Journal: Journal of Neuroinflammation

    Article Title: Histamine upregulates the expression of histamine receptors and increases the neuroprotective effect of astrocytes

    doi: 10.1186/s12974-018-1068-x

    Figure Lengend Snippet: Effects of HR antagonists on HA-induced stimulation of GDNF protein levels in astrocytes. a Incubation with histamine at the doses of 0.01 and 0.1 μg/ml for 6 and 24 h significantly promoted GDNF expression in astrocytes. b Astrocyte cells were stimulated with histamine (0.1 μg/ml) in the absence or presence of the H1R antagonist cetirizine (10 μM), the H2R antagonist ranitidine (10 μM), and the H3R antagonist carcinine (10 μM) added 30 min before stimulation. At 24 h after the addition of histamine, levels of GDNF were detected by Western blotting, quantified, and normalized to GAPDH levels. Values are expressed relative to the control, which was set to 1. ** p

    Article Snippet: Specific rabbit polyclonal anti-H1 receptor and rabbit polyclonal anti-H2 receptor antibodies were purchased from Alomone Labs Ltd. (Israel), and rabbit polyclonal anti-H4 receptor antibody was purchased from Santa Cruz (Santa Cruz Biotechnology, USA).

    Techniques: Incubation, Expressing, Western Blot

    Histamine modulates inflammation in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting (WB) and immunoreactions with anti-NADPH oxidase 2 (NOX2) (A) , anti-ARG-1 (B) . SOD1-G93A microglia stimulated with histamine (100 µM) in the presence of H1R–H2R–H3R–H4R antagonists for 6 h were subjected to immunoreactions with anti-NOX2 (C) or anti-ARG-1 (D) . SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to WB and immunoreactions with anti-CD163 (E) or anti-CD206 (F) . Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Journal: Frontiers in Immunology

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    doi: 10.3389/fimmu.2017.01689

    Figure Lengend Snippet: Histamine modulates inflammation in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting (WB) and immunoreactions with anti-NADPH oxidase 2 (NOX2) (A) , anti-ARG-1 (B) . SOD1-G93A microglia stimulated with histamine (100 µM) in the presence of H1R–H2R–H3R–H4R antagonists for 6 h were subjected to immunoreactions with anti-NOX2 (C) or anti-ARG-1 (D) . SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to WB and immunoreactions with anti-CD163 (E) or anti-CD206 (F) . Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Article Snippet: Antibodies Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

    Techniques: Western Blot

    Protein expression levels of H1-H4R in cortex and lumbar spinal cord from wild-type (WT) and SOD1-G93A mice. Cortex (A,C,E,G) and lumbar spinal cord (B,D,F,H) from WT (~20 weeks) and SOD1-G93A mice at different stages of disease (100, 140 days, and end stage) ( n = 4/group) were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to WT mice, * p

    Journal: Frontiers in Immunology

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    doi: 10.3389/fimmu.2017.01689

    Figure Lengend Snippet: Protein expression levels of H1-H4R in cortex and lumbar spinal cord from wild-type (WT) and SOD1-G93A mice. Cortex (A,C,E,G) and lumbar spinal cord (B,D,F,H) from WT (~20 weeks) and SOD1-G93A mice at different stages of disease (100, 140 days, and end stage) ( n = 4/group) were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to WT mice, * p

    Article Snippet: Antibodies Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

    Techniques: Expressing, Mouse Assay, SDS Page, Western Blot

    MAPKs are activated by histamine in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were treated with histamine (100 µM) for 15 min–1 h–6 h and equal amounts of total lysates were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-p-p38 and anti-p-38 (A) or with anti-pERK and anti-ERK (B) . (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 1 h in the presence of specific antagonists for H1R (orphenadrine, 10 µM), H2R (ranitidine, 10 µM), H3R (thioperamide, 5 µM), or H4R (JNJ-7777120, 5 µM) were subjected to immunoreactions with anti-pERK and anti-ERK. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Journal: Frontiers in Immunology

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    doi: 10.3389/fimmu.2017.01689

    Figure Lengend Snippet: MAPKs are activated by histamine in SOD1-G93A microglia. Wild-type (WT) and SOD1-G93A primary microglia were treated with histamine (100 µM) for 15 min–1 h–6 h and equal amounts of total lysates were subjected to SDS-PAGE, western blotting, and immunoreactions with anti-p-p38 and anti-p-38 (A) or with anti-pERK and anti-ERK (B) . (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 1 h in the presence of specific antagonists for H1R (orphenadrine, 10 µM), H2R (ranitidine, 10 µM), H3R (thioperamide, 5 µM), or H4R (JNJ-7777120, 5 µM) were subjected to immunoreactions with anti-pERK and anti-ERK. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Article Snippet: Antibodies Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

    Techniques: SDS Page, Western Blot

    NF-κB pathway is modulated by histamine in SOD1-G93A microglia. (A) Total RNA was extracted from SOD1-G93A microglia stimulated with histamine (100 µM) for 6–18 h and the expression profiles of IL-10, IL-6, and IL-1β were examined by qRT-PCR. (B) Wild-type (WT) and SOD1-G93A microglia exposed to 100 µM histamine for 1–6–18 h were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 6 h in the presence of H1R–H2R–H3R–H4R antagonists were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (D) SOD1-G93A microglia treated with histamine (100 µM) for 6 h in the presence of specific inhibitors of pERK (PD98059, 100 µM) and pp38 (SB203580, 20 µM) were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Journal: Frontiers in Immunology

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    doi: 10.3389/fimmu.2017.01689

    Figure Lengend Snippet: NF-κB pathway is modulated by histamine in SOD1-G93A microglia. (A) Total RNA was extracted from SOD1-G93A microglia stimulated with histamine (100 µM) for 6–18 h and the expression profiles of IL-10, IL-6, and IL-1β were examined by qRT-PCR. (B) Wild-type (WT) and SOD1-G93A microglia exposed to 100 µM histamine for 1–6–18 h were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (C) SOD1-G93A microglia stimulated with histamine (100 µM) for 6 h in the presence of H1R–H2R–H3R–H4R antagonists were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. (D) SOD1-G93A microglia treated with histamine (100 µM) for 6 h in the presence of specific inhibitors of pERK (PD98059, 100 µM) and pp38 (SB203580, 20 µM) were subjected to immunoreactions with anti-pNF-κB and anti-NF-κB. Anti-GAPDH was used for protein normalization. HA, histamine. Data represent mean ± SEM of n = 4 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Article Snippet: Antibodies Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

    Techniques: Expressing, Quantitative RT-PCR

    Histamine induces migration in SOD1-G93A microglia. (A) Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting and immunoreactions with anti-P2Y12. Anti-GAPDH was used for protein normalization. (B) SOD1-G93A microglia were stimulated with 100 µM histamine in the presence or absence of H1R–H2R–H3R–H4R antagonists or P2Y12 antagonist (MRS2395, 10 µM) for 18 h and stained with anti-CD11b (green). Scale bar 50 µm. The number of migrating cells was then quantified. (C) WT microglia were stimulated with 100 microM histamine in the presence or absence of H1R-H2R-H3R-H4R antagonists for 18 h and the number of migrating cells was quantified, HA, histamine. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Journal: Frontiers in Immunology

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    doi: 10.3389/fimmu.2017.01689

    Figure Lengend Snippet: Histamine induces migration in SOD1-G93A microglia. (A) Wild-type (WT) and SOD1-G93A primary microglia were exposed to 100 µM histamine for 1–6–18 h and equal amounts of total lysates subjected to western blotting and immunoreactions with anti-P2Y12. Anti-GAPDH was used for protein normalization. (B) SOD1-G93A microglia were stimulated with 100 µM histamine in the presence or absence of H1R–H2R–H3R–H4R antagonists or P2Y12 antagonist (MRS2395, 10 µM) for 18 h and stained with anti-CD11b (green). Scale bar 50 µm. The number of migrating cells was then quantified. (C) WT microglia were stimulated with 100 microM histamine in the presence or absence of H1R-H2R-H3R-H4R antagonists for 18 h and the number of migrating cells was quantified, HA, histamine. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by ANOVA followed by Post Hoc Tukey’s test, as referred to ctrl cells, * p

    Article Snippet: Antibodies Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

    Techniques: Migration, Western Blot, Staining

    SOD1-G93A microglia express histaminergic receptors. (A) Primary microglia from SOD1-G93A mice were stained with anti-CD11b (red) and anti-H1R, anti-H2R, anti-H3R, and anti-H4R (green). Höechst 33258 was used for nuclei (scale bar 50 µm). In inserts, wild-type (WT) microglia (scale bar 50 µm). (B) Equal amounts of WT and SOD1-G93A primary microglia total lysates were subjected to western blotting with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by Student’s t -test, as referred to WT cells, * p

    Journal: Frontiers in Immunology

    Article Title: Histamine Regulates the Inflammatory Profile of SOD1-G93A Microglia and the Histaminergic System Is Dysregulated in Amyotrophic Lateral Sclerosis

    doi: 10.3389/fimmu.2017.01689

    Figure Lengend Snippet: SOD1-G93A microglia express histaminergic receptors. (A) Primary microglia from SOD1-G93A mice were stained with anti-CD11b (red) and anti-H1R, anti-H2R, anti-H3R, and anti-H4R (green). Höechst 33258 was used for nuclei (scale bar 50 µm). In inserts, wild-type (WT) microglia (scale bar 50 µm). (B) Equal amounts of WT and SOD1-G93A primary microglia total lysates were subjected to western blotting with anti-H1R, anti-H2R, anti-H3R, and anti-H4R. Anti-GAPDH was used for protein normalization. Data represent mean ± SEM of n = 3 independent experiments. Statistical significance was calculated by Student’s t -test, as referred to WT cells, * p

    Article Snippet: Antibodies Antibodies for WB and IF were rabbit anti-HDC (1:500 WB; 1:200 IF, Abcam, USA); rabbit anti-HNMT (1:200 WB, IF, Atlas, Sweden); rabbit anti-DAO (1:200 WB and IF, Bioss, USA); rabbit anti-H1R (1:500 WB, 1:200 IF, Alomone, Israel); rabbit anti-H2R (1:500 WB and 1:200 IF, Alomone); rabbit anti-H3R (1:500 WB, 1:200 IF, Alomone); rabbit anti-H4R (1:500 WB, 1:200 IF, Santa Cruz); mouse anti-NOX2/gp91phox (1:1,000 WB, BD Transduction Laboratories, USA); rabbit anti-phospho-NF-κB p65 (Ser536) (1:500 WB, CST, USA); rabbit anti-NF-κB p65 (1:500 WB, CST); rabbit anti-phospho-p44/42 MAPK (ERK1/2) (Thr202/Tyr204) (1:1,000 WB, CST); mouse anti-p44/42 MAPK (ERK1/2) (L34F12) (1:1,000 WB, CST); rabbit anti-phospho-p38 MAPK (Thr180/Tyr182) (1:500 WB, CST); mouse anti-p38 MAPK (A-12) (1:500 WB, Santa Cruz); rabbit anti-Arginase 1 (ARG1) (1:700 WB, Abcam); rabbit anti-P2Y12 (1:200 WB, Anaspec, USA); rat anti-CD11b (1:200 IF, AbD Serotec, USA); rabbit anti-Iba1 (1:500, WB, IF, Wako, USA); rat anti-CD68 (1:500, WB, IF, AbD Serotec); rabbit anti-CD163 (1:100 WB, Santa Cruz); rabbit anti-CD206 (1:500 WB, Abcam); mouse anti-GAPDH (1:2,500 WB, Calbiochem, USA).

    Techniques: Mouse Assay, Staining, Western Blot