anti pan mglu5  (Alomone Labs)


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    Alomone Labs anti pan mglu5
    Group 1 mGluRs interact with Actn4. A , schematic of wild type Actn4 and deletion mutants fused to GST used in a pulldown assay with mGlu 1b . B , the Actn4 CH region is necessary and sufficient for mGlu 1b binding. Shown are representative immunoblots ( IB )of a pulldown assay between GST-Actn4 fusion proteins from A and lysates of HEK293 cells transfected with mGlu 1b , probed with anti-mGlu 1b , and stained with Ponceau. C and D , the mGlu 1 cytoplasmic tail is sufficient for Actn4 binding. Shown is a schematic of chimeric constructs of VSVG protein fused to mGlu 1b or mGlu 1a carboxyl tails. Yellow highlights the membrane-proximal region of the tail that is conserved between receptor isoforms (CR region, see below). Also shown are representative immunoblots of input (20 μg) and pulldown (input lysate, 2 mg) between GST-Actn4 and VSVG-mGlu 1a-Tail ( D ) and VSVG-mGlu 1b-Tail ( C ) probed with anti-mGlu 1a or anti-mGlu 1b antibody, respectively. The estimated pulldown fraction in vitro is ∼1% of input receptors. E , Actn4 interacts directly with the CR region of the mGlu 1 intracellular tail. Shown is a schematic of the CR construct and immunoblots illustrating in vitro binding of a purified His-tagged CR fragment and GST-tagged full-length Actn4 ( GST-Actn4 , left panel ) or Actn4 CH region ( GST-Actn4 1–296 , right panel ). Shown are bound and unbound (14% of input GST-tagged proteins) fractions probed with anti-GST and anti-His tag antibodies. The arrow points to full-length GST-Actn4 ( left panel ). Lower molecular mass bands were derived from partial protein fragmentation. The estimated relative bound fractions are 40% and 26% of input for GST-Actn4 and GST-Actn41–296, respectively. F , Actn4 coprecipitates with mGlu 5 in the brain. Shown are representative immunoblots of input lysate and precipitated proteins (from 2.5 mg of input lysate) probed with anti-Actn4 (input, 10 μg) and anti-mGlu 5 (input, 40 μg). The estimated immunoprecipitation (IP) is 5% and 3% of input for <t>mGlu5</t> and Actn4, respectively.
    Anti Pan Mglu5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pan mglu5/product/Alomone Labs
    Average 91 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    anti pan mglu5 - by Bioz Stars, 2022-08
    91/100 stars

    Images

    1) Product Images from "Actinin-4 Governs Dendritic Spine Dynamics and Promotes Their Remodeling by Metabotropic Glutamate Receptors *"

    Article Title: Actinin-4 Governs Dendritic Spine Dynamics and Promotes Their Remodeling by Metabotropic Glutamate Receptors *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M115.640136

    Group 1 mGluRs interact with Actn4. A , schematic of wild type Actn4 and deletion mutants fused to GST used in a pulldown assay with mGlu 1b . B , the Actn4 CH region is necessary and sufficient for mGlu 1b binding. Shown are representative immunoblots ( IB )of a pulldown assay between GST-Actn4 fusion proteins from A and lysates of HEK293 cells transfected with mGlu 1b , probed with anti-mGlu 1b , and stained with Ponceau. C and D , the mGlu 1 cytoplasmic tail is sufficient for Actn4 binding. Shown is a schematic of chimeric constructs of VSVG protein fused to mGlu 1b or mGlu 1a carboxyl tails. Yellow highlights the membrane-proximal region of the tail that is conserved between receptor isoforms (CR region, see below). Also shown are representative immunoblots of input (20 μg) and pulldown (input lysate, 2 mg) between GST-Actn4 and VSVG-mGlu 1a-Tail ( D ) and VSVG-mGlu 1b-Tail ( C ) probed with anti-mGlu 1a or anti-mGlu 1b antibody, respectively. The estimated pulldown fraction in vitro is ∼1% of input receptors. E , Actn4 interacts directly with the CR region of the mGlu 1 intracellular tail. Shown is a schematic of the CR construct and immunoblots illustrating in vitro binding of a purified His-tagged CR fragment and GST-tagged full-length Actn4 ( GST-Actn4 , left panel ) or Actn4 CH region ( GST-Actn4 1–296 , right panel ). Shown are bound and unbound (14% of input GST-tagged proteins) fractions probed with anti-GST and anti-His tag antibodies. The arrow points to full-length GST-Actn4 ( left panel ). Lower molecular mass bands were derived from partial protein fragmentation. The estimated relative bound fractions are 40% and 26% of input for GST-Actn4 and GST-Actn41–296, respectively. F , Actn4 coprecipitates with mGlu 5 in the brain. Shown are representative immunoblots of input lysate and precipitated proteins (from 2.5 mg of input lysate) probed with anti-Actn4 (input, 10 μg) and anti-mGlu 5 (input, 40 μg). The estimated immunoprecipitation (IP) is 5% and 3% of input for mGlu5 and Actn4, respectively.
    Figure Legend Snippet: Group 1 mGluRs interact with Actn4. A , schematic of wild type Actn4 and deletion mutants fused to GST used in a pulldown assay with mGlu 1b . B , the Actn4 CH region is necessary and sufficient for mGlu 1b binding. Shown are representative immunoblots ( IB )of a pulldown assay between GST-Actn4 fusion proteins from A and lysates of HEK293 cells transfected with mGlu 1b , probed with anti-mGlu 1b , and stained with Ponceau. C and D , the mGlu 1 cytoplasmic tail is sufficient for Actn4 binding. Shown is a schematic of chimeric constructs of VSVG protein fused to mGlu 1b or mGlu 1a carboxyl tails. Yellow highlights the membrane-proximal region of the tail that is conserved between receptor isoforms (CR region, see below). Also shown are representative immunoblots of input (20 μg) and pulldown (input lysate, 2 mg) between GST-Actn4 and VSVG-mGlu 1a-Tail ( D ) and VSVG-mGlu 1b-Tail ( C ) probed with anti-mGlu 1a or anti-mGlu 1b antibody, respectively. The estimated pulldown fraction in vitro is ∼1% of input receptors. E , Actn4 interacts directly with the CR region of the mGlu 1 intracellular tail. Shown is a schematic of the CR construct and immunoblots illustrating in vitro binding of a purified His-tagged CR fragment and GST-tagged full-length Actn4 ( GST-Actn4 , left panel ) or Actn4 CH region ( GST-Actn4 1–296 , right panel ). Shown are bound and unbound (14% of input GST-tagged proteins) fractions probed with anti-GST and anti-His tag antibodies. The arrow points to full-length GST-Actn4 ( left panel ). Lower molecular mass bands were derived from partial protein fragmentation. The estimated relative bound fractions are 40% and 26% of input for GST-Actn4 and GST-Actn41–296, respectively. F , Actn4 coprecipitates with mGlu 5 in the brain. Shown are representative immunoblots of input lysate and precipitated proteins (from 2.5 mg of input lysate) probed with anti-Actn4 (input, 10 μg) and anti-mGlu 5 (input, 40 μg). The estimated immunoprecipitation (IP) is 5% and 3% of input for mGlu5 and Actn4, respectively.

    Techniques Used: Binding Assay, Western Blot, Transfection, Staining, Construct, In Vitro, Purification, Derivative Assay, Immunoprecipitation

    2) Product Images from "Ablation of SNX6 leads to defects in synaptic function of CA1 pyramidal neurons and spatial memory"

    Article Title: Ablation of SNX6 leads to defects in synaptic function of CA1 pyramidal neurons and spatial memory

    Journal: eLife

    doi: 10.7554/eLife.20991

    Homer1a overexpression increases surface mGluR5 expression. This experiment serves as a control for Figure 9—figure supplement 2 and is in agreement with previous reports ( Ango et al., 2002 ). ( A ) Hippocampal neurons were co-transfected with pLL3.7.1 and EGFP-Homer1a or EGFP construct on DIV13, and fixed on DIV18. Surface mGluR5 expressions were immunostained with antibodies to mGluR5. ( B ) Quantification of surface mGluR5 signal in ( A ) (mean ± SEM, n = 30 cells, N = 3). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.035
    Figure Legend Snippet: Homer1a overexpression increases surface mGluR5 expression. This experiment serves as a control for Figure 9—figure supplement 2 and is in agreement with previous reports ( Ango et al., 2002 ). ( A ) Hippocampal neurons were co-transfected with pLL3.7.1 and EGFP-Homer1a or EGFP construct on DIV13, and fixed on DIV18. Surface mGluR5 expressions were immunostained with antibodies to mGluR5. ( B ) Quantification of surface mGluR5 signal in ( A ) (mean ± SEM, n = 30 cells, N = 3). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.035

    Techniques Used: Over Expression, Expressing, Transfection, Construct

    Ablation of SNX6 causes an increase in surface expression of mGluR5 in hippocampal neurons, which is rescued by SNX6 or Homer1c. ( A ) DIV13 neurons were transfected with constructs expressing DsRed and EGFP, EGFP-SNX6, mEmerald-Homer1c-FL or EGFP-Homer1c-C, fixed on DIV18 and immunostained with antibodies to surface-expressed mGluR5. ( B ) Quantification of surface mGluR5 fluorescence intensity per μm 2 (mean ± SEM, n = 30, N = 3, the exact p values are indicated in the bar graph). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.034
    Figure Legend Snippet: Ablation of SNX6 causes an increase in surface expression of mGluR5 in hippocampal neurons, which is rescued by SNX6 or Homer1c. ( A ) DIV13 neurons were transfected with constructs expressing DsRed and EGFP, EGFP-SNX6, mEmerald-Homer1c-FL or EGFP-Homer1c-C, fixed on DIV18 and immunostained with antibodies to surface-expressed mGluR5. ( B ) Quantification of surface mGluR5 fluorescence intensity per μm 2 (mean ± SEM, n = 30, N = 3, the exact p values are indicated in the bar graph). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.034

    Techniques Used: Expressing, Transfection, Construct, Fluorescence

    3) Product Images from "Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours"

    Article Title: Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours

    Journal: Nature biomedical engineering

    doi: 10.1038/s41551-018-0252-8

    mGluR5–CRISPR successfully promotes mGluR5 gene editing in the striatum of wild-type and Fmr1 knockout mice. a , Upper: schematic of the injection process for mGluR5–CRISPR into the striatum of wild-type (WT) and Fmr1 knockout (KO) mice. Saline or mGluR5–CRISPR was injected into the striatum (Bregma: 0.26 mm, 3 injection sites per hemisphere are indicated as blue dots, 0.4-mm interval) of WT or Fmr1 KO mice. Lower: schematic of the target sequences of Cas9 RNPs and the protospacer adjacent motif (PAM) for Grm5 knockout. b , RNA was extracted from the saline-injected control side (control) or from the mGluR5–CRISPR-injected side (mGluR5–CRISPR) of WT or Fmr1 KO mice 11 weeks after stereotaxic injections. mRNA levels of Grm5 were amplified and analysed by RT-qPCR. Fold-change of Grm5 mRNA levels are shown after normalization against PPIA mRNA levels. n = 4–6, mean ± s.e.m., *** P
    Figure Legend Snippet: mGluR5–CRISPR successfully promotes mGluR5 gene editing in the striatum of wild-type and Fmr1 knockout mice. a , Upper: schematic of the injection process for mGluR5–CRISPR into the striatum of wild-type (WT) and Fmr1 knockout (KO) mice. Saline or mGluR5–CRISPR was injected into the striatum (Bregma: 0.26 mm, 3 injection sites per hemisphere are indicated as blue dots, 0.4-mm interval) of WT or Fmr1 KO mice. Lower: schematic of the target sequences of Cas9 RNPs and the protospacer adjacent motif (PAM) for Grm5 knockout. b , RNA was extracted from the saline-injected control side (control) or from the mGluR5–CRISPR-injected side (mGluR5–CRISPR) of WT or Fmr1 KO mice 11 weeks after stereotaxic injections. mRNA levels of Grm5 were amplified and analysed by RT-qPCR. Fold-change of Grm5 mRNA levels are shown after normalization against PPIA mRNA levels. n = 4–6, mean ± s.e.m., *** P

    Techniques Used: CRISPR, Knock-Out, Mouse Assay, Injection, Amplification, Quantitative RT-PCR

    Knocking out mGluR5 using mGluR5–CRISPR significantly rescues the increased repetitive behaviours in Fmr1 knockout mice. a–c , Three weeks after stereotaxic injection of either saline (control) or mGluR5–CRISPR into the striatum of WT and Fmr1 KO mice, the marble-burying assay ( a ) or the empty cage observation test ( b , c ) was performed. a , Left: percentage of marbles buried after 30 min of the marble-burying test. Right: representative images after marble-burying assay for 30 min. Jumping ( b ) and line crossing behaviours ( c ) were scored during 10 min of an empty cage observation test. n = 10–12 for each group, mean ± s.e.m., * P
    Figure Legend Snippet: Knocking out mGluR5 using mGluR5–CRISPR significantly rescues the increased repetitive behaviours in Fmr1 knockout mice. a–c , Three weeks after stereotaxic injection of either saline (control) or mGluR5–CRISPR into the striatum of WT and Fmr1 KO mice, the marble-burying assay ( a ) or the empty cage observation test ( b , c ) was performed. a , Left: percentage of marbles buried after 30 min of the marble-burying test. Right: representative images after marble-burying assay for 30 min. Jumping ( b ) and line crossing behaviours ( c ) were scored during 10 min of an empty cage observation test. n = 10–12 for each group, mean ± s.e.m., * P

    Techniques Used: CRISPR, Knock-Out, Mouse Assay, Injection

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    • anti pan mglu5  (Alomone Labs)
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    Alomone Labs anti pan mglu5
    Group 1 mGluRs interact with Actn4. A , schematic of wild type Actn4 and deletion mutants fused to GST used in a pulldown assay with mGlu 1b . B , the Actn4 CH region is necessary and sufficient for mGlu 1b binding. Shown are representative immunoblots ( IB )of a pulldown assay between GST-Actn4 fusion proteins from A and lysates of HEK293 cells transfected with mGlu 1b , probed with anti-mGlu 1b , and stained with Ponceau. C and D , the mGlu 1 cytoplasmic tail is sufficient for Actn4 binding. Shown is a schematic of chimeric constructs of VSVG protein fused to mGlu 1b or mGlu 1a carboxyl tails. Yellow highlights the membrane-proximal region of the tail that is conserved between receptor isoforms (CR region, see below). Also shown are representative immunoblots of input (20 μg) and pulldown (input lysate, 2 mg) between GST-Actn4 and VSVG-mGlu 1a-Tail ( D ) and VSVG-mGlu 1b-Tail ( C ) probed with anti-mGlu 1a or anti-mGlu 1b antibody, respectively. The estimated pulldown fraction in vitro is ∼1% of input receptors. E , Actn4 interacts directly with the CR region of the mGlu 1 intracellular tail. Shown is a schematic of the CR construct and immunoblots illustrating in vitro binding of a purified His-tagged CR fragment and GST-tagged full-length Actn4 ( GST-Actn4 , left panel ) or Actn4 CH region ( GST-Actn4 1–296 , right panel ). Shown are bound and unbound (14% of input GST-tagged proteins) fractions probed with anti-GST and anti-His tag antibodies. The arrow points to full-length GST-Actn4 ( left panel ). Lower molecular mass bands were derived from partial protein fragmentation. The estimated relative bound fractions are 40% and 26% of input for GST-Actn4 and GST-Actn41–296, respectively. F , Actn4 coprecipitates with mGlu 5 in the brain. Shown are representative immunoblots of input lysate and precipitated proteins (from 2.5 mg of input lysate) probed with anti-Actn4 (input, 10 μg) and anti-mGlu 5 (input, 40 μg). The estimated immunoprecipitation (IP) is 5% and 3% of input for <t>mGlu5</t> and Actn4, respectively.
    Anti Pan Mglu5, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pan mglu5/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pan mglu5 - by Bioz Stars, 2022-08
    91/100 stars
    		  Buy from Supplier

    Image Search Results


    Group 1 mGluRs interact with Actn4. A , schematic of wild type Actn4 and deletion mutants fused to GST used in a pulldown assay with mGlu 1b . B , the Actn4 CH region is necessary and sufficient for mGlu 1b binding. Shown are representative immunoblots ( IB )of a pulldown assay between GST-Actn4 fusion proteins from A and lysates of HEK293 cells transfected with mGlu 1b , probed with anti-mGlu 1b , and stained with Ponceau. C and D , the mGlu 1 cytoplasmic tail is sufficient for Actn4 binding. Shown is a schematic of chimeric constructs of VSVG protein fused to mGlu 1b or mGlu 1a carboxyl tails. Yellow highlights the membrane-proximal region of the tail that is conserved between receptor isoforms (CR region, see below). Also shown are representative immunoblots of input (20 μg) and pulldown (input lysate, 2 mg) between GST-Actn4 and VSVG-mGlu 1a-Tail ( D ) and VSVG-mGlu 1b-Tail ( C ) probed with anti-mGlu 1a or anti-mGlu 1b antibody, respectively. The estimated pulldown fraction in vitro is ∼1% of input receptors. E , Actn4 interacts directly with the CR region of the mGlu 1 intracellular tail. Shown is a schematic of the CR construct and immunoblots illustrating in vitro binding of a purified His-tagged CR fragment and GST-tagged full-length Actn4 ( GST-Actn4 , left panel ) or Actn4 CH region ( GST-Actn4 1–296 , right panel ). Shown are bound and unbound (14% of input GST-tagged proteins) fractions probed with anti-GST and anti-His tag antibodies. The arrow points to full-length GST-Actn4 ( left panel ). Lower molecular mass bands were derived from partial protein fragmentation. The estimated relative bound fractions are 40% and 26% of input for GST-Actn4 and GST-Actn41–296, respectively. F , Actn4 coprecipitates with mGlu 5 in the brain. Shown are representative immunoblots of input lysate and precipitated proteins (from 2.5 mg of input lysate) probed with anti-Actn4 (input, 10 μg) and anti-mGlu 5 (input, 40 μg). The estimated immunoprecipitation (IP) is 5% and 3% of input for mGlu5 and Actn4, respectively.

    Journal: The Journal of Biological Chemistry

    Article Title: Actinin-4 Governs Dendritic Spine Dynamics and Promotes Their Remodeling by Metabotropic Glutamate Receptors *

    doi: 10.1074/jbc.M115.640136

    Figure Lengend Snippet: Group 1 mGluRs interact with Actn4. A , schematic of wild type Actn4 and deletion mutants fused to GST used in a pulldown assay with mGlu 1b . B , the Actn4 CH region is necessary and sufficient for mGlu 1b binding. Shown are representative immunoblots ( IB )of a pulldown assay between GST-Actn4 fusion proteins from A and lysates of HEK293 cells transfected with mGlu 1b , probed with anti-mGlu 1b , and stained with Ponceau. C and D , the mGlu 1 cytoplasmic tail is sufficient for Actn4 binding. Shown is a schematic of chimeric constructs of VSVG protein fused to mGlu 1b or mGlu 1a carboxyl tails. Yellow highlights the membrane-proximal region of the tail that is conserved between receptor isoforms (CR region, see below). Also shown are representative immunoblots of input (20 μg) and pulldown (input lysate, 2 mg) between GST-Actn4 and VSVG-mGlu 1a-Tail ( D ) and VSVG-mGlu 1b-Tail ( C ) probed with anti-mGlu 1a or anti-mGlu 1b antibody, respectively. The estimated pulldown fraction in vitro is ∼1% of input receptors. E , Actn4 interacts directly with the CR region of the mGlu 1 intracellular tail. Shown is a schematic of the CR construct and immunoblots illustrating in vitro binding of a purified His-tagged CR fragment and GST-tagged full-length Actn4 ( GST-Actn4 , left panel ) or Actn4 CH region ( GST-Actn4 1–296 , right panel ). Shown are bound and unbound (14% of input GST-tagged proteins) fractions probed with anti-GST and anti-His tag antibodies. The arrow points to full-length GST-Actn4 ( left panel ). Lower molecular mass bands were derived from partial protein fragmentation. The estimated relative bound fractions are 40% and 26% of input for GST-Actn4 and GST-Actn41–296, respectively. F , Actn4 coprecipitates with mGlu 5 in the brain. Shown are representative immunoblots of input lysate and precipitated proteins (from 2.5 mg of input lysate) probed with anti-Actn4 (input, 10 μg) and anti-mGlu 5 (input, 40 μg). The estimated immunoprecipitation (IP) is 5% and 3% of input for mGlu5 and Actn4, respectively.

    Article Snippet: The following antibodies were used: goat polyclonal anti-GAPDH (GenScript); chicken polyclonal anti-MAP2 (EnCor Biotech); the rabbit monoclonal antibodies anti-Actn4, anti-Actn1, and anti-mGlu5 (Epitomics); the rabbit polyclonal antibodies anti-GFP (Santa Cruz Biotechnology), anti-pan-mGlu1 , anti-pan-mGlu5 (Alomone Labs), anti-mGlu5 (GenScript), anti-PSD95 (Zymed Laboratories Inc.), anti-Actn4 (Enzo Life Sciences), anti-phospho-ERK1/2(Thr-202/Tyr-204), anti-ERK1/2, anti-phospho-CaMKII(Thr-286/Thr-287), and anti-CaMKII (Cell Signaling Technology); and the mouse monoclonal antibodies anti-MAP2 clone AP20 (Roche Applied Science), anti-actinin clone EA-53 (Sigma-Aldrich), anti-Actn4 (Abnova), anti-mGlu1a (BD Biosciences), anti-PSD95 clone K28/43 (University of California Davis/National Institutes of Health NeuroMab Facility), and anti-His tag (Aviva Systems Biology).

    Techniques: Binding Assay, Western Blot, Transfection, Staining, Construct, In Vitro, Purification, Derivative Assay, Immunoprecipitation

    Homer1a overexpression increases surface mGluR5 expression. This experiment serves as a control for Figure 9—figure supplement 2 and is in agreement with previous reports ( Ango et al., 2002 ). ( A ) Hippocampal neurons were co-transfected with pLL3.7.1 and EGFP-Homer1a or EGFP construct on DIV13, and fixed on DIV18. Surface mGluR5 expressions were immunostained with antibodies to mGluR5. ( B ) Quantification of surface mGluR5 signal in ( A ) (mean ± SEM, n = 30 cells, N = 3). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.035

    Journal: eLife

    Article Title: Ablation of SNX6 leads to defects in synaptic function of CA1 pyramidal neurons and spatial memory

    doi: 10.7554/eLife.20991

    Figure Lengend Snippet: Homer1a overexpression increases surface mGluR5 expression. This experiment serves as a control for Figure 9—figure supplement 2 and is in agreement with previous reports ( Ango et al., 2002 ). ( A ) Hippocampal neurons were co-transfected with pLL3.7.1 and EGFP-Homer1a or EGFP construct on DIV13, and fixed on DIV18. Surface mGluR5 expressions were immunostained with antibodies to mGluR5. ( B ) Quantification of surface mGluR5 signal in ( A ) (mean ± SEM, n = 30 cells, N = 3). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.035

    Article Snippet: Antibodies Antibodies used in this study are: mouse anti-GluA1 (MAB2263), mouse anti-GluA2 (MAB397), mouse anti-Tau1 (MAB3420), and mouse anti-MAP2 (MAB3418) from Millipore (Billerica, MA); mouse anti-SYP (D-4) (sc-17750), mouse anti-PSD95 (6G6) (sc-32291), mouse anti-DIC (74-1) (sc-13524), goat anti-Homer1a (M-13) (sc-8922), rabbit anti-Homer1b/c (H-174) (sc-20807), mouse anti-SNX6 (D-5) (sc-365965), goat anti-SNX6 (N-19) (sc-8679), and rabbit anti-Rab5B (A-20) (sc-598) from Santa Cruz Biotechnology (Santa Cruz, CA); mouse anti-p150Glued (610474), mouse anti-EEA1 (610457) from BD Biosciences (San Diego, CA); rabbit anti-clathrin (ab21679), rabbit anti-TGN46 (ab50595), rat anti-CTIP2 (ab18465), rabbit anti-SNX1 (ab134126) and rabbit anti-Rab4 (ab109009) from Abcam; rabbit anti-GluN1 (D65B7) from Cell Signaling Technology (Mississauga, ON, Canada); rabbit anti-GluN2B (AGC-003) and rabbit anti-mGluR5 (AGC-007) from Alomone labs (Jerusalem, Israel); rabbit and mouse anti-GFP (MBL598, D153-3), rabbit and mouse anti-RFP (PM005, M155-3) from Medical and Biological Laboratories (Naka-kuNagoya, Japan); mouse anti-β-actin (A5441) (Sigma-Aldrich, St. Louis, MO); mouse anti-Golgi97 (A21270) from Invitrogen (Carlsbad, CA); goat anti-VPS35 (PAB7499) from Abnova (Taipei, Taiwan) and rabbit anti-GluN2A (612–401-D89) from Rockland Immunochemicals (Limerick, PA); rabbit anti-Rab7 (#9367) from Cell Signaling Technology (Mississauga, ON, Canada); rabbit anti-Rab11 (3H18L, 700184) from Life Technologies (Carlsbad, CA); Rabbit anti-SNX6 was described previously ( ).

    Techniques: Over Expression, Expressing, Transfection, Construct

    Ablation of SNX6 causes an increase in surface expression of mGluR5 in hippocampal neurons, which is rescued by SNX6 or Homer1c. ( A ) DIV13 neurons were transfected with constructs expressing DsRed and EGFP, EGFP-SNX6, mEmerald-Homer1c-FL or EGFP-Homer1c-C, fixed on DIV18 and immunostained with antibodies to surface-expressed mGluR5. ( B ) Quantification of surface mGluR5 fluorescence intensity per μm 2 (mean ± SEM, n = 30, N = 3, the exact p values are indicated in the bar graph). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.034

    Journal: eLife

    Article Title: Ablation of SNX6 leads to defects in synaptic function of CA1 pyramidal neurons and spatial memory

    doi: 10.7554/eLife.20991

    Figure Lengend Snippet: Ablation of SNX6 causes an increase in surface expression of mGluR5 in hippocampal neurons, which is rescued by SNX6 or Homer1c. ( A ) DIV13 neurons were transfected with constructs expressing DsRed and EGFP, EGFP-SNX6, mEmerald-Homer1c-FL or EGFP-Homer1c-C, fixed on DIV18 and immunostained with antibodies to surface-expressed mGluR5. ( B ) Quantification of surface mGluR5 fluorescence intensity per μm 2 (mean ± SEM, n = 30, N = 3, the exact p values are indicated in the bar graph). Bar: 2 μm. DOI: http://dx.doi.org/10.7554/eLife.20991.034

    Article Snippet: Antibodies Antibodies used in this study are: mouse anti-GluA1 (MAB2263), mouse anti-GluA2 (MAB397), mouse anti-Tau1 (MAB3420), and mouse anti-MAP2 (MAB3418) from Millipore (Billerica, MA); mouse anti-SYP (D-4) (sc-17750), mouse anti-PSD95 (6G6) (sc-32291), mouse anti-DIC (74-1) (sc-13524), goat anti-Homer1a (M-13) (sc-8922), rabbit anti-Homer1b/c (H-174) (sc-20807), mouse anti-SNX6 (D-5) (sc-365965), goat anti-SNX6 (N-19) (sc-8679), and rabbit anti-Rab5B (A-20) (sc-598) from Santa Cruz Biotechnology (Santa Cruz, CA); mouse anti-p150Glued (610474), mouse anti-EEA1 (610457) from BD Biosciences (San Diego, CA); rabbit anti-clathrin (ab21679), rabbit anti-TGN46 (ab50595), rat anti-CTIP2 (ab18465), rabbit anti-SNX1 (ab134126) and rabbit anti-Rab4 (ab109009) from Abcam; rabbit anti-GluN1 (D65B7) from Cell Signaling Technology (Mississauga, ON, Canada); rabbit anti-GluN2B (AGC-003) and rabbit anti-mGluR5 (AGC-007) from Alomone labs (Jerusalem, Israel); rabbit and mouse anti-GFP (MBL598, D153-3), rabbit and mouse anti-RFP (PM005, M155-3) from Medical and Biological Laboratories (Naka-kuNagoya, Japan); mouse anti-β-actin (A5441) (Sigma-Aldrich, St. Louis, MO); mouse anti-Golgi97 (A21270) from Invitrogen (Carlsbad, CA); goat anti-VPS35 (PAB7499) from Abnova (Taipei, Taiwan) and rabbit anti-GluN2A (612–401-D89) from Rockland Immunochemicals (Limerick, PA); rabbit anti-Rab7 (#9367) from Cell Signaling Technology (Mississauga, ON, Canada); rabbit anti-Rab11 (3H18L, 700184) from Life Technologies (Carlsbad, CA); Rabbit anti-SNX6 was described previously ( ).

    Techniques: Expressing, Transfection, Construct, Fluorescence

    mGluR5–CRISPR successfully promotes mGluR5 gene editing in the striatum of wild-type and Fmr1 knockout mice. a , Upper: schematic of the injection process for mGluR5–CRISPR into the striatum of wild-type (WT) and Fmr1 knockout (KO) mice. Saline or mGluR5–CRISPR was injected into the striatum (Bregma: 0.26 mm, 3 injection sites per hemisphere are indicated as blue dots, 0.4-mm interval) of WT or Fmr1 KO mice. Lower: schematic of the target sequences of Cas9 RNPs and the protospacer adjacent motif (PAM) for Grm5 knockout. b , RNA was extracted from the saline-injected control side (control) or from the mGluR5–CRISPR-injected side (mGluR5–CRISPR) of WT or Fmr1 KO mice 11 weeks after stereotaxic injections. mRNA levels of Grm5 were amplified and analysed by RT-qPCR. Fold-change of Grm5 mRNA levels are shown after normalization against PPIA mRNA levels. n = 4–6, mean ± s.e.m., *** P

    Journal: Nature biomedical engineering

    Article Title: Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours

    doi: 10.1038/s41551-018-0252-8

    Figure Lengend Snippet: mGluR5–CRISPR successfully promotes mGluR5 gene editing in the striatum of wild-type and Fmr1 knockout mice. a , Upper: schematic of the injection process for mGluR5–CRISPR into the striatum of wild-type (WT) and Fmr1 knockout (KO) mice. Saline or mGluR5–CRISPR was injected into the striatum (Bregma: 0.26 mm, 3 injection sites per hemisphere are indicated as blue dots, 0.4-mm interval) of WT or Fmr1 KO mice. Lower: schematic of the target sequences of Cas9 RNPs and the protospacer adjacent motif (PAM) for Grm5 knockout. b , RNA was extracted from the saline-injected control side (control) or from the mGluR5–CRISPR-injected side (mGluR5–CRISPR) of WT or Fmr1 KO mice 11 weeks after stereotaxic injections. mRNA levels of Grm5 were amplified and analysed by RT-qPCR. Fold-change of Grm5 mRNA levels are shown after normalization against PPIA mRNA levels. n = 4–6, mean ± s.e.m., *** P

    Article Snippet: The mouse monoclonal RFP antibody (6G6) was purchased from ChromoTek, the chicken polyclonal GFP antibody (GFP-1020) from Aves Labs (Tigard), the rabbit polyclonal GFAP antibody (AB5804) and the mouse monoclonal NeuN antibody (MAB377) from Millipore, the rabbit polyclonal IBA1 antibody (019–19741) from Wako Chemicals, and the rabbit polyclonal mGluR5 antibody (AGC-007) was from Alomone Labs.

    Techniques: CRISPR, Knock-Out, Mouse Assay, Injection, Amplification, Quantitative RT-PCR

    Knocking out mGluR5 using mGluR5–CRISPR significantly rescues the increased repetitive behaviours in Fmr1 knockout mice. a–c , Three weeks after stereotaxic injection of either saline (control) or mGluR5–CRISPR into the striatum of WT and Fmr1 KO mice, the marble-burying assay ( a ) or the empty cage observation test ( b , c ) was performed. a , Left: percentage of marbles buried after 30 min of the marble-burying test. Right: representative images after marble-burying assay for 30 min. Jumping ( b ) and line crossing behaviours ( c ) were scored during 10 min of an empty cage observation test. n = 10–12 for each group, mean ± s.e.m., * P

    Journal: Nature biomedical engineering

    Article Title: Nanoparticle delivery of CRISPR into the brain rescues a mouse model of fragile X syndrome from exaggerated repetitive behaviours

    doi: 10.1038/s41551-018-0252-8

    Figure Lengend Snippet: Knocking out mGluR5 using mGluR5–CRISPR significantly rescues the increased repetitive behaviours in Fmr1 knockout mice. a–c , Three weeks after stereotaxic injection of either saline (control) or mGluR5–CRISPR into the striatum of WT and Fmr1 KO mice, the marble-burying assay ( a ) or the empty cage observation test ( b , c ) was performed. a , Left: percentage of marbles buried after 30 min of the marble-burying test. Right: representative images after marble-burying assay for 30 min. Jumping ( b ) and line crossing behaviours ( c ) were scored during 10 min of an empty cage observation test. n = 10–12 for each group, mean ± s.e.m., * P

    Article Snippet: The mouse monoclonal RFP antibody (6G6) was purchased from ChromoTek, the chicken polyclonal GFP antibody (GFP-1020) from Aves Labs (Tigard), the rabbit polyclonal GFAP antibody (AB5804) and the mouse monoclonal NeuN antibody (MAB377) from Millipore, the rabbit polyclonal IBA1 antibody (019–19741) from Wako Chemicals, and the rabbit polyclonal mGluR5 antibody (AGC-007) was from Alomone Labs.

    Techniques: CRISPR, Knock-Out, Mouse Assay, Injection