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glua1 cell signaling (13185) antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc glua1 cell signaling (13185) antibody
    Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and <t>GluA1</t> (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.
    Glua1 Cell Signaling (13185) Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glua1 cell signaling (13185) antibody/product/Cell Signaling Technology Inc
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    Images

    1) Product Images from "Establishment of a Depression Model Using Dexamethasone-treated Three-dimensional Cultured Rat Cortical Cells"

    Article Title: Establishment of a Depression Model Using Dexamethasone-treated Three-dimensional Cultured Rat Cortical Cells

    Journal: Clinical Psychopharmacology and Neuroscience

    doi: 10.9758/cpn.25.1269

    Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and GluA1 (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.
    Figure Legend Snippet: Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and GluA1 (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.

    Techniques Used: Phospho-proteomics, Expressing, Western Blot, Immunofluorescence, Marker, In Vitro, Binding Assay

    Schematic diagram showing the molecular mechanisms underlying DEX-induced impaired neuroplasticity. Exposure to DEX downregulates signaling pathways that affect neuroplasticity, including BDNF, sirtuin 1, and mTORC1 signaling. Activation of the mTORC1 signaling pathway induces the synthesis of synaptic proteins as well as BDNF. Secreted BDNF interacts with its receptor TrkB, further activating mTORC1 signaling via PI3K/Akt and MEK/ERK1/2. Therefore, synaptic plasticity is enhanced. Sirtuin 1 is also involved in the regulation of neuroplasticity. The original illustration was created using BioRender (biorender.com). Akt, protein kinase B; AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid; BDNF, brain-derived neurotrophic factor; DEX, dexamethasone; ERK1/2, extracellular signal-regulated kinase 1/2; GluA1, AMPA receptor subunit glutamate receptor 1; MEK, mitogen‑activated protein kinase; mTORC1, mechanistic target of rapamycin complex I; p70S6K, p70S6 kinase; PI3K, phosphatidyl inositol-3 kinase; PSD-95, postsynaptic density protein-95; TrKB, tropomyosin receptor kinase B; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1.
    Figure Legend Snippet: Schematic diagram showing the molecular mechanisms underlying DEX-induced impaired neuroplasticity. Exposure to DEX downregulates signaling pathways that affect neuroplasticity, including BDNF, sirtuin 1, and mTORC1 signaling. Activation of the mTORC1 signaling pathway induces the synthesis of synaptic proteins as well as BDNF. Secreted BDNF interacts with its receptor TrkB, further activating mTORC1 signaling via PI3K/Akt and MEK/ERK1/2. Therefore, synaptic plasticity is enhanced. Sirtuin 1 is also involved in the regulation of neuroplasticity. The original illustration was created using BioRender (biorender.com). Akt, protein kinase B; AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid; BDNF, brain-derived neurotrophic factor; DEX, dexamethasone; ERK1/2, extracellular signal-regulated kinase 1/2; GluA1, AMPA receptor subunit glutamate receptor 1; MEK, mitogen‑activated protein kinase; mTORC1, mechanistic target of rapamycin complex I; p70S6K, p70S6 kinase; PI3K, phosphatidyl inositol-3 kinase; PSD-95, postsynaptic density protein-95; TrKB, tropomyosin receptor kinase B; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1.

    Techniques Used: Protein-Protein interactions, Activation Assay, Derivative Assay, Binding Assay



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    Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and <t>GluA1</t> (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.
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    Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and <t>GluA1</t> (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.
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    Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and <t>GluA1</t> (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.
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    Figure 1. LRRK2 RC mutation increases synaptic glutamate receptor content in the striatum. (A) Schematic diagram of LRRK2 protein highlighting the armadillo repeats (ARM), ankyrin (ANK) repeats, Ras of complex (ROC), C-terminal of ROC (COR), kin (KIN), and WD40 domains. Knock- in mice expressing the R1441C and G2019S mutations found in the ROC and kinase domains respectively, crossed with either Drd1-Tomato or Drd2-eGFP mouse lines. (B) Workflow schematic for subcellular fractionation of striatal homogenate for the enrichment of postsynaptic density fraction (PSD). (C) Representative western blot analysis of the subcellular fractionation results, showing supernatant (S1), crude synaptosomal preparation (P2), PSD, and Triton soluble fractions (TSF). (D) Western blot analysis of +/+, +/RC, and +/GS P2 striatal fractions probed for p-PKA substrates, pS845 <t>GluA1,</t> total GluA1, pT72Rab8A, total Rab8A, pT73Rab10, total Rab10, and PSD95. (E) Western blot analysis of +/+, +/RC, and +/GS mice probed for GluA1, p-PKA, and PSD95. S1 and PSD fractions are shown. (F-G) Quantification of GluA1 Figure 1 continued on next page
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    Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and GluA1 (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.

    Journal: Clinical Psychopharmacology and Neuroscience

    Article Title: Establishment of a Depression Model Using Dexamethasone-treated Three-dimensional Cultured Rat Cortical Cells

    doi: 10.9758/cpn.25.1269

    Figure Lengend Snippet: Changes in the phosphorylation of mTORC1-mediated proteins and in the expression of synaptic proteins after DEX exposure. Beginning on DIV 10, the neural spheroids were exposed to 100 μM DEX for 5 days. Western blotting and immunofluorescence were performed using each primary antibody (n = 4 biological replicates). (A) Western blotting revealed the levels of phospho-Ser 2448 -mTORC1 (a), phospho- Thr 37/46 -4E-BP1 (b), and phospho-Thr 389 -p70S6K (c). (B) Western blot analysis and representative images of immunoblots of PSD-95 (a) and GluA1 (b) are shown. Synaptic markers: PSD-95 (green) costained with the neuronal marker MAP-2 (red); GluA1 (green) costained with MAP-2 (red). Scale bar: 100 μm. The data are presented as the means ± standard deviations. mTORC1, mechanistic target of rapamycin complex I; DEX, dexamethasone; DIV, days in vitro; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1; PSD-95, postsynaptic density protein-95; GluA1, AMPA receptor subunit glutamate receptor 1; MAP-2, microtubule-associated protein-2; CON, controls. * p < 0.05 or ** p < 0.01 vs. CON.

    Article Snippet: GluA1 , Cell Signaling Technology (13185) , AB_2732897 , IF (1:1,000) WB (1:1,000).

    Techniques: Phospho-proteomics, Expressing, Western Blot, Immunofluorescence, Marker, In Vitro, Binding Assay

    Schematic diagram showing the molecular mechanisms underlying DEX-induced impaired neuroplasticity. Exposure to DEX downregulates signaling pathways that affect neuroplasticity, including BDNF, sirtuin 1, and mTORC1 signaling. Activation of the mTORC1 signaling pathway induces the synthesis of synaptic proteins as well as BDNF. Secreted BDNF interacts with its receptor TrkB, further activating mTORC1 signaling via PI3K/Akt and MEK/ERK1/2. Therefore, synaptic plasticity is enhanced. Sirtuin 1 is also involved in the regulation of neuroplasticity. The original illustration was created using BioRender (biorender.com). Akt, protein kinase B; AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid; BDNF, brain-derived neurotrophic factor; DEX, dexamethasone; ERK1/2, extracellular signal-regulated kinase 1/2; GluA1, AMPA receptor subunit glutamate receptor 1; MEK, mitogen‑activated protein kinase; mTORC1, mechanistic target of rapamycin complex I; p70S6K, p70S6 kinase; PI3K, phosphatidyl inositol-3 kinase; PSD-95, postsynaptic density protein-95; TrKB, tropomyosin receptor kinase B; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1.

    Journal: Clinical Psychopharmacology and Neuroscience

    Article Title: Establishment of a Depression Model Using Dexamethasone-treated Three-dimensional Cultured Rat Cortical Cells

    doi: 10.9758/cpn.25.1269

    Figure Lengend Snippet: Schematic diagram showing the molecular mechanisms underlying DEX-induced impaired neuroplasticity. Exposure to DEX downregulates signaling pathways that affect neuroplasticity, including BDNF, sirtuin 1, and mTORC1 signaling. Activation of the mTORC1 signaling pathway induces the synthesis of synaptic proteins as well as BDNF. Secreted BDNF interacts with its receptor TrkB, further activating mTORC1 signaling via PI3K/Akt and MEK/ERK1/2. Therefore, synaptic plasticity is enhanced. Sirtuin 1 is also involved in the regulation of neuroplasticity. The original illustration was created using BioRender (biorender.com). Akt, protein kinase B; AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid; BDNF, brain-derived neurotrophic factor; DEX, dexamethasone; ERK1/2, extracellular signal-regulated kinase 1/2; GluA1, AMPA receptor subunit glutamate receptor 1; MEK, mitogen‑activated protein kinase; mTORC1, mechanistic target of rapamycin complex I; p70S6K, p70S6 kinase; PI3K, phosphatidyl inositol-3 kinase; PSD-95, postsynaptic density protein-95; TrKB, tropomyosin receptor kinase B; 4E-BP1, eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1.

    Article Snippet: GluA1 , Cell Signaling Technology (13185) , AB_2732897 , IF (1:1,000) WB (1:1,000).

    Techniques: Protein-Protein interactions, Activation Assay, Derivative Assay, Binding Assay

    Figure 1. LRRK2 RC mutation increases synaptic glutamate receptor content in the striatum. (A) Schematic diagram of LRRK2 protein highlighting the armadillo repeats (ARM), ankyrin (ANK) repeats, Ras of complex (ROC), C-terminal of ROC (COR), kin (KIN), and WD40 domains. Knock- in mice expressing the R1441C and G2019S mutations found in the ROC and kinase domains respectively, crossed with either Drd1-Tomato or Drd2-eGFP mouse lines. (B) Workflow schematic for subcellular fractionation of striatal homogenate for the enrichment of postsynaptic density fraction (PSD). (C) Representative western blot analysis of the subcellular fractionation results, showing supernatant (S1), crude synaptosomal preparation (P2), PSD, and Triton soluble fractions (TSF). (D) Western blot analysis of +/+, +/RC, and +/GS P2 striatal fractions probed for p-PKA substrates, pS845 GluA1, total GluA1, pT72Rab8A, total Rab8A, pT73Rab10, total Rab10, and PSD95. (E) Western blot analysis of +/+, +/RC, and +/GS mice probed for GluA1, p-PKA, and PSD95. S1 and PSD fractions are shown. (F-G) Quantification of GluA1 Figure 1 continued on next page

    Journal: eLife

    Article Title: Pathway-specific dysregulation of striatal excitatory synapses by LRRK2 mutations

    doi: 10.7554/elife.58997

    Figure Lengend Snippet: Figure 1. LRRK2 RC mutation increases synaptic glutamate receptor content in the striatum. (A) Schematic diagram of LRRK2 protein highlighting the armadillo repeats (ARM), ankyrin (ANK) repeats, Ras of complex (ROC), C-terminal of ROC (COR), kin (KIN), and WD40 domains. Knock- in mice expressing the R1441C and G2019S mutations found in the ROC and kinase domains respectively, crossed with either Drd1-Tomato or Drd2-eGFP mouse lines. (B) Workflow schematic for subcellular fractionation of striatal homogenate for the enrichment of postsynaptic density fraction (PSD). (C) Representative western blot analysis of the subcellular fractionation results, showing supernatant (S1), crude synaptosomal preparation (P2), PSD, and Triton soluble fractions (TSF). (D) Western blot analysis of +/+, +/RC, and +/GS P2 striatal fractions probed for p-PKA substrates, pS845 GluA1, total GluA1, pT72Rab8A, total Rab8A, pT73Rab10, total Rab10, and PSD95. (E) Western blot analysis of +/+, +/RC, and +/GS mice probed for GluA1, p-PKA, and PSD95. S1 and PSD fractions are shown. (F-G) Quantification of GluA1 Figure 1 continued on next page

    Article Snippet: (species) or resource Designation Source or reference Identifiers Additional information Antibody Phospho-PKA Substrate (RRXS*/T*) (100G7E) rabbit monoclonal Cell Signaling Technology RRID:AB_331817 WB (1:1000) Antibody AMPA Receptor 1 (GluA1) (D4N9V) rabbit monoclonal Cell Signaling Technology RRID:AB_2732897 WB (1:1000) Antibody Phospho-AMPA Receptor 1 (GluA1) (Ser845) (D10G5) rabbit monoclonal Cell Signaling Technology RRID:AB_10860773 WB (1:1000) Antibody Synaptophysin rabbit polyclonal Cell Signaling Technology RRID:AB_1904154 WB (1:1000) Antibody Anti-RAB8A antibody [EPR14873] rabbit monoclonal Abcam RRID:AB_2814989 WB (1:1000) Antibody RAB8A (phospho T72) [MJF-R20] rabbit monoclonal Abcam RRID:AB_2814988 WB (1:1000) Antibody Rab10 (D36C4) XP Rabbit mAb rabbit monoclonal Cell signaling technology RRID:AB_10828219 WB (1:1000) Antibody Anti-RAB10 (phospho T73) antibody [MJF-R21] rabbit monoclonal Abcam RRID:AB_2811274 WB (1:1000) Antibody HOMER1 polyclonal antibody rabbit polyclonal Proteintech RRID:AB_2295573 WB (1:1000) Antibody GluA1/GluR1 glutamate receptor clone N355/1 mouse monoclonal UC Davis/NIH NeuroMab Facility RRID:AB_2315840 IF (1:300) Antibody PSD-95 monoclonal (6G6-1C9) mouse monoclonal Invitrogen RRID:AB_325399 WB (1:1000) IF (1:300) Antibody PSD-95 polyclonal rabbit polyclonal Invitrogen RRID:AB_87705 IF (1:300) Antibody GFP chicken polyclonal Invitrogen RRID:AB_2534023 IF (1:1000) Antibody mCherry (16D7) rat monoclonal Invitrogen RRID:AB_2536611 IF (1:1000) Antibody Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP Invitrogen RRID:AB_2533947 WB (1:5000) Antibody Goat anti-Rabbit IgG (H+L) Secondary Antibody, HRP Invitrogen RRID:AB_2533967 WB (1:5000) Antibody Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 647 Invitrogen RRID:AB_162542 IF (1:300) Antibody Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 568 Invitrogen RRID:AB_2534017 IF (1:300) Antibody Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 568 Invitrogen RRID:AB_2534121 IF (1:300)

    Techniques: Mutagenesis, Knock-In, Expressing, Fractionation, Western Blot