Review



rabbit anti-syntaxin-12/13  (Synaptic Systems)


Bioz Verified Symbol Synaptic Systems is a verified supplier
Bioz Manufacturer Symbol Synaptic Systems manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 90
      Buy from Supplier

    Structured Review

    Synaptic Systems rabbit anti-syntaxin-12/13
    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="250" height="auto" />
    Rabbit Anti Syntaxin 12/13, supplied by Synaptic Systems, 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/rabbit anti-syntaxin-12/13/product/Synaptic Systems
    Average 90 stars, based on 1 article reviews
    rabbit anti-syntaxin-12/13 - by Bioz Stars, 2026-05
    90/100 stars

    Images

    1) Product Images from "Proteomic Analysis Reveals the Composition of Glutamatergic Organelles of Auditory Inner Hair Cells"

    Article Title: Proteomic Analysis Reveals the Composition of Glutamatergic Organelles of Auditory Inner Hair Cells

    Journal: Molecular & Cellular Proteomics : MCP

    doi: 10.1016/j.mcpro.2023.100704

    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " title="... displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle.

    Techniques Used: Expressing, Protein Enrichment, Marker, Binding Assay, Mass Spectrometry

    Comparative analysis of enriched proteins in P8 and P23 VGluT3 immunoisolates revealed a small overlap in protein identifications between ages and a different relative enrichment for shared proteins . A , Venn diagram showing the overlap between the significantly enriched proteins (>1.5-fold enrichment) at P8 and P23. B and C , ranking of significantly enriched proteins according to their relative enrichment in VGluT3 over Control IgG immunoisolates and input S2, before ( B ) and after ( C ) hearing onset. Relative Enrichment corresponds to the average of log 2 iBAQ (VGluT3/Input) and log 2 iBAQ (VGluT3/Control IgG). Displayed are only enriched proteins shown also in <xref ref-type=Figure 4 , D and E ( upper right quadrant and >1.5-fold enrichment). Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). Source data are available for this figure ( supplemental Table S3 ). " title="... enrichment). Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Comparative analysis of enriched proteins in P8 and P23 VGluT3 immunoisolates revealed a small overlap in protein identifications between ages and a different relative enrichment for shared proteins . A , Venn diagram showing the overlap between the significantly enriched proteins (>1.5-fold enrichment) at P8 and P23. B and C , ranking of significantly enriched proteins according to their relative enrichment in VGluT3 over Control IgG immunoisolates and input S2, before ( B ) and after ( C ) hearing onset. Relative Enrichment corresponds to the average of log 2 iBAQ (VGluT3/Input) and log 2 iBAQ (VGluT3/Control IgG). Displayed are only enriched proteins shown also in Figure 4 , D and E ( upper right quadrant and >1.5-fold enrichment). Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). Source data are available for this figure ( supplemental Table S3 ).

    Techniques Used:

    Functional annotation of mature VGluT3-associated IHC proteome reveals a mixed SV-endosomal signature . A , sunburst diagram with functional annotation of the enriched proteins in VGluT3 immunoisolates at P23. Proteins were grouped for display according to their cellular component and their involvement in trafficking events. Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular component and biological function. For detailed annotations see <xref ref-type=supplemental Table S4 . B–F , Scatter plots showing positively enriched proteins in VGluT3 immunoisolates at P23 when compared to both control IgG and input; log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted. Displayed are proteins involved in trafficking events in different trafficking organelles (SV, endolysosomal, Golgi, and ER proteins) including SNAREs and resident proteins. Several Rab GTPase proteins, mostly of endolysosomal nature, were also enriched ( F ). Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular compartment and biological function. For detailed annotation see supplemental Table S4 . In ( B – F ) gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community but is still annotated in most databases as syntaxin-12). For ranking of proteins according to their relative enrichment, see supplemental Fig. S5 . Source data are available for this figure ( supplemental Table S2 ). ER, endoplasmic reticulum; PM, plasma membrane; SV, synaptic vesicle. " title="... displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Functional annotation of mature VGluT3-associated IHC proteome reveals a mixed SV-endosomal signature . A , sunburst diagram with functional annotation of the enriched proteins in VGluT3 immunoisolates at P23. Proteins were grouped for display according to their cellular component and their involvement in trafficking events. Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular component and biological function. For detailed annotations see supplemental Table S4 . B–F , Scatter plots showing positively enriched proteins in VGluT3 immunoisolates at P23 when compared to both control IgG and input; log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted. Displayed are proteins involved in trafficking events in different trafficking organelles (SV, endolysosomal, Golgi, and ER proteins) including SNAREs and resident proteins. Several Rab GTPase proteins, mostly of endolysosomal nature, were also enriched ( F ). Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular compartment and biological function. For detailed annotation see supplemental Table S4 . In ( B – F ) gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community but is still annotated in most databases as syntaxin-12). For ranking of proteins according to their relative enrichment, see supplemental Fig. S5 . Source data are available for this figure ( supplemental Table S2 ). ER, endoplasmic reticulum; PM, plasma membrane; SV, synaptic vesicle.

    Techniques Used: Functional Assay, Membrane

    Immunolocalization analysis of SNARE, SV, and kinase proteins in the adult organ of Corti . A–D , VAMP-7 ( A ), syntaxin-12/13 ( B ), syntaxin-8 ( C ), and syntaxin-7 ( D ), all highly enriched in our MS experiments, are expressed in IHCs. E–H , some classical neuronal SV proteins SCAMP1 ( E ), V-ATPase ( F ), and SV2B ( G ), and the kinase PKCα ( H ), all enriched in our MS experiments, are expressed in IHCs. Note that all proteins localize to the basolateral region of the IHCs, although in some cases the protein is also expressed in supporting cells and/or in afferent (postsynaptic) and efferent fibers of SGNs. Images correspond to high magnification views of representative P15–25 IHCs immunolabeled with antibodies against the candidate proteins ( red ), the ribbon marker CtBP2/RIBEYE ( green ), and the IHC marker otoferlin ( blue ). The upper panels show overviews of representative IHCs, displaying maximum intensity projections of 5 to 10 confocal optical sections through the longitudinal axis of the IHCs (scale bars: 5 μm). The bottom panels show a zoom into the synaptic area, displaying single confocal optical sections through the longitudinal axis of a single IHCs at the basal region (scale bars: 2 μm). IHC, inner hair cell; MS, mass spectrometry; SGN, spiral ganglion neuron.
    Figure Legend Snippet: Immunolocalization analysis of SNARE, SV, and kinase proteins in the adult organ of Corti . A–D , VAMP-7 ( A ), syntaxin-12/13 ( B ), syntaxin-8 ( C ), and syntaxin-7 ( D ), all highly enriched in our MS experiments, are expressed in IHCs. E–H , some classical neuronal SV proteins SCAMP1 ( E ), V-ATPase ( F ), and SV2B ( G ), and the kinase PKCα ( H ), all enriched in our MS experiments, are expressed in IHCs. Note that all proteins localize to the basolateral region of the IHCs, although in some cases the protein is also expressed in supporting cells and/or in afferent (postsynaptic) and efferent fibers of SGNs. Images correspond to high magnification views of representative P15–25 IHCs immunolabeled with antibodies against the candidate proteins ( red ), the ribbon marker CtBP2/RIBEYE ( green ), and the IHC marker otoferlin ( blue ). The upper panels show overviews of representative IHCs, displaying maximum intensity projections of 5 to 10 confocal optical sections through the longitudinal axis of the IHCs (scale bars: 5 μm). The bottom panels show a zoom into the synaptic area, displaying single confocal optical sections through the longitudinal axis of a single IHCs at the basal region (scale bars: 2 μm). IHC, inner hair cell; MS, mass spectrometry; SGN, spiral ganglion neuron.

    Techniques Used: Immunolabeling, Marker, Mass Spectrometry



    Similar Products

    syntaxin 13  (Developmental Studies Hybridoma Bank)
    95
    Developmental Studies Hybridoma Bank syntaxin 13
    Syntaxin 13, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/syntaxin 13/product/Developmental Studies Hybridoma Bank
    Average 95 stars, based on 1 article reviews
    syntaxin 13 - by Bioz Stars, 2026-05
    95/100 stars
    		  Buy from Supplier
    syntaxin 13  (Bioss)
    90
    Bioss syntaxin 13
    Syntaxin 13, supplied by Bioss, 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/syntaxin 13/product/Bioss
    Average 90 stars, based on 1 article reviews
    syntaxin 13 - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    rabbit anti-syntaxin-12/13  (Synaptic Systems)
    90
    Synaptic Systems rabbit anti-syntaxin-12/13
    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="250" height="auto" />
    Rabbit Anti Syntaxin 12/13, supplied by Synaptic Systems, 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/rabbit anti-syntaxin-12/13/product/Synaptic Systems
    Average 90 stars, based on 1 article reviews
    rabbit anti-syntaxin-12/13 - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    anti-syntaxin 13  (Stressgen Biotechnologies)
    90
    Stressgen Biotechnologies anti-syntaxin 13
    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="250" height="auto" />
    Anti Syntaxin 13, supplied by Stressgen Biotechnologies, 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/anti-syntaxin 13/product/Stressgen Biotechnologies
    Average 90 stars, based on 1 article reviews
    anti-syntaxin 13 - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    anti-syntaxin 13  (Synaptic Systems)
    90
    Synaptic Systems anti-syntaxin 13
    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="250" height="auto" />
    Anti Syntaxin 13, supplied by Synaptic Systems, 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/anti-syntaxin 13/product/Synaptic Systems
    Average 90 stars, based on 1 article reviews
    anti-syntaxin 13 - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    mouse monoclonal anti-(syntaxin 13) antibody  (Stressgen Biotechnologies)
    90
    Stressgen Biotechnologies mouse monoclonal anti-(syntaxin 13) antibody
    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="250" height="auto" />
    Mouse Monoclonal Anti (Syntaxin 13) Antibody, supplied by Stressgen Biotechnologies, 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/mouse monoclonal anti-(syntaxin 13) antibody/product/Stressgen Biotechnologies
    Average 90 stars, based on 1 article reviews
    mouse monoclonal anti-(syntaxin 13) antibody - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    mouse monoclonal anti-syntaxin-12/13 (stx-13)  (Synaptic Systems)
    90
    Synaptic Systems mouse monoclonal anti-syntaxin-12/13 (stx-13)
    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="250" height="auto" />
    Mouse Monoclonal Anti Syntaxin 12/13 (Stx 13), supplied by Synaptic Systems, 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/mouse monoclonal anti-syntaxin-12/13 (stx-13)/product/Synaptic Systems
    Average 90 stars, based on 1 article reviews
    mouse monoclonal anti-syntaxin-12/13 (stx-13) - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    mouse monoclonal anti-syntaxin-12/13 (stx-13  (Synaptic Systems)
    90
    Synaptic Systems mouse monoclonal anti-syntaxin-12/13 (stx-13
    NHE6 colocalizes with early and recycling endosomal markers at dendritic spines of mouse primary hippocampal neurons. A, C–E, Representative confocal micrographs show immunofluorescent localization of NHE6 and its colocalization with early and recycling endosomes positive for EEA1, AF-Tfn, or <t>Stx-13</t> in mouse primary hippocampal neurons (14+ DIV). Arrowheads denote protein localization at different regions of dendritic spines. A, NHE6 localizes to the somatodendritic compartment of an mGFP-positive neuron. It also localizes to neighboring unlabeled neurons and to cells of the surrounding glial cell bed. Scale bar, 20 μm. B, Simplified schematic representation of the endosomal recycling pathway at the dendritic spine surface. C, NHE6-EEA1 colocalization occurs at a subset of NHE6-positive dendritic spines in primary hippocampal neurons. EEA1 is almost exclusively found in close proximity to or partially overlapping NHE6 puncta. Magnification in all parts (C–E) is identical. Scale bar, 2 μm. D, mGFP-positive dendrite following incubation with AF-Tfn (100 μg/ml) for 1 h at 37°C to label recycling endosomes. Puncta of internalized AF-Tfn are frequently adjacent to or overlapping NHE6. E, NHE6-Stx-13 colocalization is seen only at a subset of NHE6-positive dendritic spines in primary hippocampal neurons. Stx-13 is almost exclusively found in close proximity or partially overlapping NHE6 puncta. F, Quantitative summary of NHE6 colocalization with recycling endosomal markers in dendritic spines of primary hippocampal neurons. NHE6 localizes to 90% of mature dendritic spines, showing colocalization with EEA1 at 35% of spines, AF-Tfn at 50% of spines, and with Stx-13 at 40% of spines. By the Mander's coefficient, the majority of Stx-13 and EEA1-positive puncta are significantly overlapped by NHE6 (Stx-13-NHE6: 0.8564, p < 0.05; EEA1-NHE6: 0.9611, p < 0.05), with approximately half of AF-Tfn overlapping with NHE6 (AF-Tfn-NHE6: 0.5733, p < 0.05). NHE6 colocalization with recycling endosomes occurs primarily at the spine base (a) and/or head (c), and to a minimal extent at the spine neck (b) (see inset schematic). Note: All immunopositive spines are described as a/b/c. Percentage values for dendritic spine areas (i.e., a, b, and c) do not add up to 100%, as individual spines could fall into more than one category. n = 163 dendritic spines from 172.78 μm dendrite from six neurons, nAF-Tfn = 221 dendritic spines from 228.70 μm dendrite from six neurons, 266 dendritic spines from 323.53 μm dendrite from eight neurons. Mean ± SEM localization per dendrite segment. White dashed line denotes secondary dendrite of an mGFP-positive dendrite. EnV, Endocytic vesicle; EE, early endosome; RE, recycling endosome; ExV, exocytic vesicle; EEA1, early endosome antigen 1; AF-Tfn, Alexa Fluor 568-conjugated transferrin; Stx-13, syntaxin-13; mGFP, membrane-tagged enhanced green fluorescent protein.
    Mouse Monoclonal Anti Syntaxin 12/13 (Stx 13, supplied by Synaptic Systems, 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/mouse monoclonal anti-syntaxin-12/13 (stx-13/product/Synaptic Systems
    Average 90 stars, based on 1 article reviews
    mouse monoclonal anti-syntaxin-12/13 (stx-13 - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier
    rabbit anti-syntaxin 13  (Synaptic Systems)
    90
    Synaptic Systems rabbit anti-syntaxin 13
    Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for <t>syntaxin</t> <t>13</t> and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.
    Rabbit Anti Syntaxin 13, supplied by Synaptic Systems, 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/rabbit anti-syntaxin 13/product/Synaptic Systems
    Average 90 stars, based on 1 article reviews
    rabbit anti-syntaxin 13 - by Bioz Stars, 2026-05
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in <xref ref-type=Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle. " width="100%" height="100%">

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Proteomic Analysis Reveals the Composition of Glutamatergic Organelles of Auditory Inner Hair Cells

    doi: 10.1016/j.mcpro.2023.100704

    Figure Lengend Snippet: MS-based comparative enrichment analysis of VGluT3 immunoisolates’ proteome reveals age-dependent changes in the expression of synaptic and trafficking proteins . A , schematic representation of the immunoisolation approach to isolate IHC VGluT3-positive vesicular structures from immature (P8) and mature (P23) organs of Corti. S2 fraction was prepared as described in Figure 1 from ∼100 organs of Corti (per biological replicate) and used as starting material in VGluT3-and control IgG-specific immunoisolations. Two technical replicates (indicated as T1 and T2) from two independent immunoisolation procedures (biological replicates R1 and R2) were measured. B and C , approach used for the analysis of the MS data. B , protein enrichment was assessed by comparing VGluT3 immunoisolates with both control IgG immunoisolates and input S2 samples. The –log 10 adjusted p value was plotted against the log 2 iBAQ fold change of VGluT3 over control (IgG or input), with a significant t test FDR threshold of 5% and S 0 = 0. C , to visualize protein enrichment in VGluT3 immunoisolates as compared to both control IgG and input S2, log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted; proteins in the upper right quadrant were enriched in VGluT3 immunoisolates – a threshold of log 2 iBAQ fold difference >0.7 was set to consider only proteins with at least 1.5-fold enrichment. D and E , Scatter plots showing differential enrichment of proteins in VGluT3 immunoisolates when compared to both control IgG and inputs at P8 ( D ) and P23 ( E ); displayed are IHC marker proteins VGluT3 and otoferlin, classical SV proteins, SNAREs, SNARE-binding proteins and other proteins. Numbers in parenthesis refer to the total number of significantly enriched proteins in VGluT3 immunoisolates over control IgG and Input (>1.5-fold enrichment). Gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). See also supplemental Fig. S3 for individual volcano plots. Source data are available for this figure ( supplemental Table S2 ). IHC, inner hair cell; MS, mass spectrometry; SV, synaptic vesicle.

    Article Snippet: The following primary antibodies were used: rabbit anti-VAMP-7 (232003, Synaptic Systems), rabbit anti-syntaxin-6 (110062, Synaptic Systems), rabbit anti-syntaxin-7 (110072, Synaptic Systems), rabbit anti-syntaxin-8 (110083, Synaptic Systems), rabbit anti-syntaxin-12/13 (110133, Synaptic Systems), rabbit anti-syntaxin-16 (110162, Synaptic Systems), rabbit anti-SCAMP1 (PA1-739, Thermo Fisher Scientific), mouse anti-V-ATPase (149011, Synaptic Systems), mouse anti-SV2B (119111, Synaptic Systems), rabbit anti-Vti1A (165002, Synaptic Systems), rabbit anti-PKC alpha [Y124] (ab32376, Abcam), rabbit anti-YKT6 (ab236583, Abcam), rabbit anti-VAP-A (249002, Synaptic Systems), mouse anti-VCP (MA3-004, Thermo Fisher Scientific), mouse anti-otoferlin [13A9] (ab53233, Abcam), rabbit anti-otoferlin (178003, Synaptic Systems), rabbit anti-VGluT3 (135203, Synaptic Systems), guinea pig anti-VGluT3 (135204, Synaptic Systems), mouse anti-synapsin-1 (106001, Synaptic Systems), goat IgG anti-CtBP2 [E−16] (sc-5967, Santa Cruz Biotechnology), rabbit anti-Munc18-1 (116002, Synaptic Systems), rabbit anti-Munc18-2 (116102, Synaptic Systems), rabbit anti-Munc18-3 (116202, Synaptic Systems), and mouse anti-VAMP-2 (104211, Synaptic Systems).

    Techniques: Expressing, Protein Enrichment, Marker, Binding Assay, Mass Spectrometry

    Comparative analysis of enriched proteins in P8 and P23 VGluT3 immunoisolates revealed a small overlap in protein identifications between ages and a different relative enrichment for shared proteins . A , Venn diagram showing the overlap between the significantly enriched proteins (>1.5-fold enrichment) at P8 and P23. B and C , ranking of significantly enriched proteins according to their relative enrichment in VGluT3 over Control IgG immunoisolates and input S2, before ( B ) and after ( C ) hearing onset. Relative Enrichment corresponds to the average of log 2 iBAQ (VGluT3/Input) and log 2 iBAQ (VGluT3/Control IgG). Displayed are only enriched proteins shown also in <xref ref-type=Figure 4 , D and E ( upper right quadrant and >1.5-fold enrichment). Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). Source data are available for this figure ( supplemental Table S3 ). " width="100%" height="100%">

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Proteomic Analysis Reveals the Composition of Glutamatergic Organelles of Auditory Inner Hair Cells

    doi: 10.1016/j.mcpro.2023.100704

    Figure Lengend Snippet: Comparative analysis of enriched proteins in P8 and P23 VGluT3 immunoisolates revealed a small overlap in protein identifications between ages and a different relative enrichment for shared proteins . A , Venn diagram showing the overlap between the significantly enriched proteins (>1.5-fold enrichment) at P8 and P23. B and C , ranking of significantly enriched proteins according to their relative enrichment in VGluT3 over Control IgG immunoisolates and input S2, before ( B ) and after ( C ) hearing onset. Relative Enrichment corresponds to the average of log 2 iBAQ (VGluT3/Input) and log 2 iBAQ (VGluT3/Control IgG). Displayed are only enriched proteins shown also in Figure 4 , D and E ( upper right quadrant and >1.5-fold enrichment). Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community, but is still annotated in most databases as syntaxin-12). Source data are available for this figure ( supplemental Table S3 ).

    Article Snippet: The following primary antibodies were used: rabbit anti-VAMP-7 (232003, Synaptic Systems), rabbit anti-syntaxin-6 (110062, Synaptic Systems), rabbit anti-syntaxin-7 (110072, Synaptic Systems), rabbit anti-syntaxin-8 (110083, Synaptic Systems), rabbit anti-syntaxin-12/13 (110133, Synaptic Systems), rabbit anti-syntaxin-16 (110162, Synaptic Systems), rabbit anti-SCAMP1 (PA1-739, Thermo Fisher Scientific), mouse anti-V-ATPase (149011, Synaptic Systems), mouse anti-SV2B (119111, Synaptic Systems), rabbit anti-Vti1A (165002, Synaptic Systems), rabbit anti-PKC alpha [Y124] (ab32376, Abcam), rabbit anti-YKT6 (ab236583, Abcam), rabbit anti-VAP-A (249002, Synaptic Systems), mouse anti-VCP (MA3-004, Thermo Fisher Scientific), mouse anti-otoferlin [13A9] (ab53233, Abcam), rabbit anti-otoferlin (178003, Synaptic Systems), rabbit anti-VGluT3 (135203, Synaptic Systems), guinea pig anti-VGluT3 (135204, Synaptic Systems), mouse anti-synapsin-1 (106001, Synaptic Systems), goat IgG anti-CtBP2 [E−16] (sc-5967, Santa Cruz Biotechnology), rabbit anti-Munc18-1 (116002, Synaptic Systems), rabbit anti-Munc18-2 (116102, Synaptic Systems), rabbit anti-Munc18-3 (116202, Synaptic Systems), and mouse anti-VAMP-2 (104211, Synaptic Systems).

    Techniques:

    Functional annotation of mature VGluT3-associated IHC proteome reveals a mixed SV-endosomal signature . A , sunburst diagram with functional annotation of the enriched proteins in VGluT3 immunoisolates at P23. Proteins were grouped for display according to their cellular component and their involvement in trafficking events. Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular component and biological function. For detailed annotations see <xref ref-type=supplemental Table S4 . B–F , Scatter plots showing positively enriched proteins in VGluT3 immunoisolates at P23 when compared to both control IgG and input; log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted. Displayed are proteins involved in trafficking events in different trafficking organelles (SV, endolysosomal, Golgi, and ER proteins) including SNAREs and resident proteins. Several Rab GTPase proteins, mostly of endolysosomal nature, were also enriched ( F ). Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular compartment and biological function. For detailed annotation see supplemental Table S4 . In ( B – F ) gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community but is still annotated in most databases as syntaxin-12). For ranking of proteins according to their relative enrichment, see supplemental Fig. S5 . Source data are available for this figure ( supplemental Table S2 ). ER, endoplasmic reticulum; PM, plasma membrane; SV, synaptic vesicle. " width="100%" height="100%">

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Proteomic Analysis Reveals the Composition of Glutamatergic Organelles of Auditory Inner Hair Cells

    doi: 10.1016/j.mcpro.2023.100704

    Figure Lengend Snippet: Functional annotation of mature VGluT3-associated IHC proteome reveals a mixed SV-endosomal signature . A , sunburst diagram with functional annotation of the enriched proteins in VGluT3 immunoisolates at P23. Proteins were grouped for display according to their cellular component and their involvement in trafficking events. Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular component and biological function. For detailed annotations see supplemental Table S4 . B–F , Scatter plots showing positively enriched proteins in VGluT3 immunoisolates at P23 when compared to both control IgG and input; log 2 iBAQ fold change VGluT3/Control IgG vs. log 2 iBAQ fold change VGluT3/Input was plotted. Displayed are proteins involved in trafficking events in different trafficking organelles (SV, endolysosomal, Golgi, and ER proteins) including SNAREs and resident proteins. Several Rab GTPase proteins, mostly of endolysosomal nature, were also enriched ( F ). Annotations were done manually and based on information available in several databases; proteins were grouped according to cellular compartment and biological function. For detailed annotation see supplemental Table S4 . In ( B – F ) gene names are displayed. Stx12 gene annotated in UniProt refers to syntaxin-12/13 protein (syntaxin-12 and syntaxin-13 are the same protein; syntaxin-13 is the accepted term by the scientific community but is still annotated in most databases as syntaxin-12). For ranking of proteins according to their relative enrichment, see supplemental Fig. S5 . Source data are available for this figure ( supplemental Table S2 ). ER, endoplasmic reticulum; PM, plasma membrane; SV, synaptic vesicle.

    Article Snippet: The following primary antibodies were used: rabbit anti-VAMP-7 (232003, Synaptic Systems), rabbit anti-syntaxin-6 (110062, Synaptic Systems), rabbit anti-syntaxin-7 (110072, Synaptic Systems), rabbit anti-syntaxin-8 (110083, Synaptic Systems), rabbit anti-syntaxin-12/13 (110133, Synaptic Systems), rabbit anti-syntaxin-16 (110162, Synaptic Systems), rabbit anti-SCAMP1 (PA1-739, Thermo Fisher Scientific), mouse anti-V-ATPase (149011, Synaptic Systems), mouse anti-SV2B (119111, Synaptic Systems), rabbit anti-Vti1A (165002, Synaptic Systems), rabbit anti-PKC alpha [Y124] (ab32376, Abcam), rabbit anti-YKT6 (ab236583, Abcam), rabbit anti-VAP-A (249002, Synaptic Systems), mouse anti-VCP (MA3-004, Thermo Fisher Scientific), mouse anti-otoferlin [13A9] (ab53233, Abcam), rabbit anti-otoferlin (178003, Synaptic Systems), rabbit anti-VGluT3 (135203, Synaptic Systems), guinea pig anti-VGluT3 (135204, Synaptic Systems), mouse anti-synapsin-1 (106001, Synaptic Systems), goat IgG anti-CtBP2 [E−16] (sc-5967, Santa Cruz Biotechnology), rabbit anti-Munc18-1 (116002, Synaptic Systems), rabbit anti-Munc18-2 (116102, Synaptic Systems), rabbit anti-Munc18-3 (116202, Synaptic Systems), and mouse anti-VAMP-2 (104211, Synaptic Systems).

    Techniques: Functional Assay, Membrane

    Immunolocalization analysis of SNARE, SV, and kinase proteins in the adult organ of Corti . A–D , VAMP-7 ( A ), syntaxin-12/13 ( B ), syntaxin-8 ( C ), and syntaxin-7 ( D ), all highly enriched in our MS experiments, are expressed in IHCs. E–H , some classical neuronal SV proteins SCAMP1 ( E ), V-ATPase ( F ), and SV2B ( G ), and the kinase PKCα ( H ), all enriched in our MS experiments, are expressed in IHCs. Note that all proteins localize to the basolateral region of the IHCs, although in some cases the protein is also expressed in supporting cells and/or in afferent (postsynaptic) and efferent fibers of SGNs. Images correspond to high magnification views of representative P15–25 IHCs immunolabeled with antibodies against the candidate proteins ( red ), the ribbon marker CtBP2/RIBEYE ( green ), and the IHC marker otoferlin ( blue ). The upper panels show overviews of representative IHCs, displaying maximum intensity projections of 5 to 10 confocal optical sections through the longitudinal axis of the IHCs (scale bars: 5 μm). The bottom panels show a zoom into the synaptic area, displaying single confocal optical sections through the longitudinal axis of a single IHCs at the basal region (scale bars: 2 μm). IHC, inner hair cell; MS, mass spectrometry; SGN, spiral ganglion neuron.

    Journal: Molecular & Cellular Proteomics : MCP

    Article Title: Proteomic Analysis Reveals the Composition of Glutamatergic Organelles of Auditory Inner Hair Cells

    doi: 10.1016/j.mcpro.2023.100704

    Figure Lengend Snippet: Immunolocalization analysis of SNARE, SV, and kinase proteins in the adult organ of Corti . A–D , VAMP-7 ( A ), syntaxin-12/13 ( B ), syntaxin-8 ( C ), and syntaxin-7 ( D ), all highly enriched in our MS experiments, are expressed in IHCs. E–H , some classical neuronal SV proteins SCAMP1 ( E ), V-ATPase ( F ), and SV2B ( G ), and the kinase PKCα ( H ), all enriched in our MS experiments, are expressed in IHCs. Note that all proteins localize to the basolateral region of the IHCs, although in some cases the protein is also expressed in supporting cells and/or in afferent (postsynaptic) and efferent fibers of SGNs. Images correspond to high magnification views of representative P15–25 IHCs immunolabeled with antibodies against the candidate proteins ( red ), the ribbon marker CtBP2/RIBEYE ( green ), and the IHC marker otoferlin ( blue ). The upper panels show overviews of representative IHCs, displaying maximum intensity projections of 5 to 10 confocal optical sections through the longitudinal axis of the IHCs (scale bars: 5 μm). The bottom panels show a zoom into the synaptic area, displaying single confocal optical sections through the longitudinal axis of a single IHCs at the basal region (scale bars: 2 μm). IHC, inner hair cell; MS, mass spectrometry; SGN, spiral ganglion neuron.

    Article Snippet: The following primary antibodies were used: rabbit anti-VAMP-7 (232003, Synaptic Systems), rabbit anti-syntaxin-6 (110062, Synaptic Systems), rabbit anti-syntaxin-7 (110072, Synaptic Systems), rabbit anti-syntaxin-8 (110083, Synaptic Systems), rabbit anti-syntaxin-12/13 (110133, Synaptic Systems), rabbit anti-syntaxin-16 (110162, Synaptic Systems), rabbit anti-SCAMP1 (PA1-739, Thermo Fisher Scientific), mouse anti-V-ATPase (149011, Synaptic Systems), mouse anti-SV2B (119111, Synaptic Systems), rabbit anti-Vti1A (165002, Synaptic Systems), rabbit anti-PKC alpha [Y124] (ab32376, Abcam), rabbit anti-YKT6 (ab236583, Abcam), rabbit anti-VAP-A (249002, Synaptic Systems), mouse anti-VCP (MA3-004, Thermo Fisher Scientific), mouse anti-otoferlin [13A9] (ab53233, Abcam), rabbit anti-otoferlin (178003, Synaptic Systems), rabbit anti-VGluT3 (135203, Synaptic Systems), guinea pig anti-VGluT3 (135204, Synaptic Systems), mouse anti-synapsin-1 (106001, Synaptic Systems), goat IgG anti-CtBP2 [E−16] (sc-5967, Santa Cruz Biotechnology), rabbit anti-Munc18-1 (116002, Synaptic Systems), rabbit anti-Munc18-2 (116102, Synaptic Systems), rabbit anti-Munc18-3 (116202, Synaptic Systems), and mouse anti-VAMP-2 (104211, Synaptic Systems).

    Techniques: Immunolabeling, Marker, Mass Spectrometry

    NHE6 colocalizes with early and recycling endosomal markers at dendritic spines of mouse primary hippocampal neurons. A, C–E, Representative confocal micrographs show immunofluorescent localization of NHE6 and its colocalization with early and recycling endosomes positive for EEA1, AF-Tfn, or Stx-13 in mouse primary hippocampal neurons (14+ DIV). Arrowheads denote protein localization at different regions of dendritic spines. A, NHE6 localizes to the somatodendritic compartment of an mGFP-positive neuron. It also localizes to neighboring unlabeled neurons and to cells of the surrounding glial cell bed. Scale bar, 20 μm. B, Simplified schematic representation of the endosomal recycling pathway at the dendritic spine surface. C, NHE6-EEA1 colocalization occurs at a subset of NHE6-positive dendritic spines in primary hippocampal neurons. EEA1 is almost exclusively found in close proximity to or partially overlapping NHE6 puncta. Magnification in all parts (C–E) is identical. Scale bar, 2 μm. D, mGFP-positive dendrite following incubation with AF-Tfn (100 μg/ml) for 1 h at 37°C to label recycling endosomes. Puncta of internalized AF-Tfn are frequently adjacent to or overlapping NHE6. E, NHE6-Stx-13 colocalization is seen only at a subset of NHE6-positive dendritic spines in primary hippocampal neurons. Stx-13 is almost exclusively found in close proximity or partially overlapping NHE6 puncta. F, Quantitative summary of NHE6 colocalization with recycling endosomal markers in dendritic spines of primary hippocampal neurons. NHE6 localizes to 90% of mature dendritic spines, showing colocalization with EEA1 at 35% of spines, AF-Tfn at 50% of spines, and with Stx-13 at 40% of spines. By the Mander's coefficient, the majority of Stx-13 and EEA1-positive puncta are significantly overlapped by NHE6 (Stx-13-NHE6: 0.8564, p < 0.05; EEA1-NHE6: 0.9611, p < 0.05), with approximately half of AF-Tfn overlapping with NHE6 (AF-Tfn-NHE6: 0.5733, p < 0.05). NHE6 colocalization with recycling endosomes occurs primarily at the spine base (a) and/or head (c), and to a minimal extent at the spine neck (b) (see inset schematic). Note: All immunopositive spines are described as a/b/c. Percentage values for dendritic spine areas (i.e., a, b, and c) do not add up to 100%, as individual spines could fall into more than one category. n = 163 dendritic spines from 172.78 μm dendrite from six neurons, nAF-Tfn = 221 dendritic spines from 228.70 μm dendrite from six neurons, 266 dendritic spines from 323.53 μm dendrite from eight neurons. Mean ± SEM localization per dendrite segment. White dashed line denotes secondary dendrite of an mGFP-positive dendrite. EnV, Endocytic vesicle; EE, early endosome; RE, recycling endosome; ExV, exocytic vesicle; EEA1, early endosome antigen 1; AF-Tfn, Alexa Fluor 568-conjugated transferrin; Stx-13, syntaxin-13; mGFP, membrane-tagged enhanced green fluorescent protein.

    Journal: The Journal of Neuroscience

    Article Title: Enhanced Recruitment of Endosomal Na + /H + Exchanger NHE6 into Dendritic Spines of Hippocampal Pyramidal Neurons during NMDA Receptor-Dependent Long-Term Potentiation

    doi: 10.1523/JNEUROSCI.2583-12.2013

    Figure Lengend Snippet: NHE6 colocalizes with early and recycling endosomal markers at dendritic spines of mouse primary hippocampal neurons. A, C–E, Representative confocal micrographs show immunofluorescent localization of NHE6 and its colocalization with early and recycling endosomes positive for EEA1, AF-Tfn, or Stx-13 in mouse primary hippocampal neurons (14+ DIV). Arrowheads denote protein localization at different regions of dendritic spines. A, NHE6 localizes to the somatodendritic compartment of an mGFP-positive neuron. It also localizes to neighboring unlabeled neurons and to cells of the surrounding glial cell bed. Scale bar, 20 μm. B, Simplified schematic representation of the endosomal recycling pathway at the dendritic spine surface. C, NHE6-EEA1 colocalization occurs at a subset of NHE6-positive dendritic spines in primary hippocampal neurons. EEA1 is almost exclusively found in close proximity to or partially overlapping NHE6 puncta. Magnification in all parts (C–E) is identical. Scale bar, 2 μm. D, mGFP-positive dendrite following incubation with AF-Tfn (100 μg/ml) for 1 h at 37°C to label recycling endosomes. Puncta of internalized AF-Tfn are frequently adjacent to or overlapping NHE6. E, NHE6-Stx-13 colocalization is seen only at a subset of NHE6-positive dendritic spines in primary hippocampal neurons. Stx-13 is almost exclusively found in close proximity or partially overlapping NHE6 puncta. F, Quantitative summary of NHE6 colocalization with recycling endosomal markers in dendritic spines of primary hippocampal neurons. NHE6 localizes to 90% of mature dendritic spines, showing colocalization with EEA1 at 35% of spines, AF-Tfn at 50% of spines, and with Stx-13 at 40% of spines. By the Mander's coefficient, the majority of Stx-13 and EEA1-positive puncta are significantly overlapped by NHE6 (Stx-13-NHE6: 0.8564, p < 0.05; EEA1-NHE6: 0.9611, p < 0.05), with approximately half of AF-Tfn overlapping with NHE6 (AF-Tfn-NHE6: 0.5733, p < 0.05). NHE6 colocalization with recycling endosomes occurs primarily at the spine base (a) and/or head (c), and to a minimal extent at the spine neck (b) (see inset schematic). Note: All immunopositive spines are described as a/b/c. Percentage values for dendritic spine areas (i.e., a, b, and c) do not add up to 100%, as individual spines could fall into more than one category. n = 163 dendritic spines from 172.78 μm dendrite from six neurons, nAF-Tfn = 221 dendritic spines from 228.70 μm dendrite from six neurons, 266 dendritic spines from 323.53 μm dendrite from eight neurons. Mean ± SEM localization per dendrite segment. White dashed line denotes secondary dendrite of an mGFP-positive dendrite. EnV, Endocytic vesicle; EE, early endosome; RE, recycling endosome; ExV, exocytic vesicle; EEA1, early endosome antigen 1; AF-Tfn, Alexa Fluor 568-conjugated transferrin; Stx-13, syntaxin-13; mGFP, membrane-tagged enhanced green fluorescent protein.

    Article Snippet: Protein localization studies using immunofluorescence and immunoblotting were performed using the following commercial antibodies: mouse monoclonal anti-hemagglutinin (HA) antibody (Covance), rabbit polyclonal anti-HA (Abcam), anti-β-tubulin antibody (Sigma-Aldrich), anti-transferrin receptor antibody (Invitrogen), mouse monoclonal anti-syntaxin-12/13 (Stx-13; Synaptic Systems), mouse monoclonal early endosomal antigen 1 (EEA1; Sigma), and mouse monoclonal anti-GluA1 (GluA1/GluR1; Synaptic Systems).

    Techniques: Incubation

    Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.

    Journal: PLoS Biology

    Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

    doi: 10.1371/journal.pbio.1000283

    Figure Lengend Snippet: Immunogold EM of hippocampal neurons labeled with 10 nm protein A gold for Rab4 and with 15 nm protein A gold for GRASP-1 (A), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for GRASP-1 (B), with 10 nm protein A gold for syntaxin 13 and with 15 nm protein A gold for Rab4 (C), or with 15 nm protein A gold for GRASP-1, with 5 nm protein gold for syntaxin 13, and with 10 nm protein A gold for rab4 (D). Arrow denotes tubular endosomal membrane to which GRASP-1, syntaxin 13, and Rab4 localized. EE indicates early endosomes and scale bar is 100 nm.

    Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

    Techniques: Labeling

    (A) Lysates of COS-7 cells cotransfected with GFP-GRASP-1 and myc-syntaxins were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. (B) Lysates of COS-7 cells cotransfected with GFP-syntaxin 13 and full-length myc-GRASP-1 (1–837) or truncated myc-GRASP-1 constructs (1–695 or 695–837) were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. Asterisk indicates background band. Arrows point to co-precipitated GRASP-1 proteins. (C) Binding assay using lysates of COS-7 cells expressing myc-syntaxin 13 with or without GFP-GRASP-1 and GMP-PNP-charged GST-rab4. Note that myc-syntaxin 13 is only isolated on the beads in the presence of GRASP-1. (D) Binding assay using lysate of COS-7 cells transfected with GFP-GRASP-1(594–837) and GST-syntaxins without transmembrane domain (ΔTM). GRASP-1 was analyzed by Western blot with antibody against GFP. (E) Binding assay of 35 S-labeled GRASP-1 and immobilized GST-syntaxin 13ΔTM.

    Journal: PLoS Biology

    Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

    doi: 10.1371/journal.pbio.1000283

    Figure Lengend Snippet: (A) Lysates of COS-7 cells cotransfected with GFP-GRASP-1 and myc-syntaxins were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. (B) Lysates of COS-7 cells cotransfected with GFP-syntaxin 13 and full-length myc-GRASP-1 (1–837) or truncated myc-GRASP-1 constructs (1–695 or 695–837) were immunoprecipitated with anti-GFP antibody and analyzed by Western blot. Asterisk indicates background band. Arrows point to co-precipitated GRASP-1 proteins. (C) Binding assay using lysates of COS-7 cells expressing myc-syntaxin 13 with or without GFP-GRASP-1 and GMP-PNP-charged GST-rab4. Note that myc-syntaxin 13 is only isolated on the beads in the presence of GRASP-1. (D) Binding assay using lysate of COS-7 cells transfected with GFP-GRASP-1(594–837) and GST-syntaxins without transmembrane domain (ΔTM). GRASP-1 was analyzed by Western blot with antibody against GFP. (E) Binding assay of 35 S-labeled GRASP-1 and immobilized GST-syntaxin 13ΔTM.

    Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

    Techniques: Immunoprecipitation, Western Blot, Construct, Binding Assay, Expressing, Isolation, Transfection, Labeling

    (A) Representative image of hippocampal neuron triple transfected at DIV13 for 4 d with GFP-Rab4, HA-GRASP-1, and myc-syntaxin 13 and labeled with anti-HA (blue) or anti-myc (red) antibodies. Magnified region of the cell body is shown to indicate the strong colocalization of GRASP-1, Rab4, and syntaxin 13. (B) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-syntaxin 13 (red). (C) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-Neep21 (red). (D) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with myc-syntaxin 13 and control vector or HA-GRASP-1 and labeled with anti-myc (green), anti-HA (blue), and anti-Neep21 (red). (E) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with GFP-Rab4, HA-Rab11, and control vector or myc-syntaxin 13ΔTM and labeled with anti-myc (blue) and anti-HA (red). (F) Percentage of colocalization between HA-GRASP-1 and myc-syntaxin 1 or myc-syntaxin 13 in neurons. (G) Percentage of colocalization between myc-syntaxin 13 and Neep21 in dendrites as indicated in (D). (H) Percentage of colocalization between GFP-Rab4 and HA-Rab11 domains in dendrites expressing myc-syntaxin 13ΔTM as indicated in (E). Error bars indicate S.E.M. ** p <0.005. *** p <0.0005. Bar in A is 10 µm; Bar in (B–E) is 1 µm.

    Journal: PLoS Biology

    Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

    doi: 10.1371/journal.pbio.1000283

    Figure Lengend Snippet: (A) Representative image of hippocampal neuron triple transfected at DIV13 for 4 d with GFP-Rab4, HA-GRASP-1, and myc-syntaxin 13 and labeled with anti-HA (blue) or anti-myc (red) antibodies. Magnified region of the cell body is shown to indicate the strong colocalization of GRASP-1, Rab4, and syntaxin 13. (B) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-syntaxin 13 (red). (C) Representative images of dendrites of hippocampal neurons transfected at DIV13 with GFP-GRASP-1 for 4 d and labeled with anti-Neep21 (red). (D) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with myc-syntaxin 13 and control vector or HA-GRASP-1 and labeled with anti-myc (green), anti-HA (blue), and anti-Neep21 (red). (E) Representative images of dendrites of hippocampal neurons cotransfected at DIV13 for 4 d with GFP-Rab4, HA-Rab11, and control vector or myc-syntaxin 13ΔTM and labeled with anti-myc (blue) and anti-HA (red). (F) Percentage of colocalization between HA-GRASP-1 and myc-syntaxin 1 or myc-syntaxin 13 in neurons. (G) Percentage of colocalization between myc-syntaxin 13 and Neep21 in dendrites as indicated in (D). (H) Percentage of colocalization between GFP-Rab4 and HA-Rab11 domains in dendrites expressing myc-syntaxin 13ΔTM as indicated in (E). Error bars indicate S.E.M. ** p <0.005. *** p <0.0005. Bar in A is 10 µm; Bar in (B–E) is 1 µm.

    Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

    Techniques: Transfection, Labeling, Plasmid Preparation, Expressing

    Endosomes can be viewed as mosaic distribution of Rab4, Rab5, and Rab11 domains that dynamically interact via effector proteins and SNAREs. The Rab5 domain allows entry into the early/sorting endosome, whereas the Rab4 and Rab11 domains contain the machinery that is necessary for sorting and recycling membranes and receptors back to the plasma membrane. (A) GRASP-1 binds to Rab4 and syntaxin 13 and couples Rab4 and Rab11 recycling endosomes. The complex formed between GRASP-1 and t-SNARE syntaxin 13 might mediate fusion between Rab4 and Rab11 endosomes. (B) Absence of GRASP-1 interferes with complex formation at the recycling step, causing cargo accumulation in early endosomes, impairment of receptor expression, and changes in spine morphology. (C) Overexpression of GRASP-1 leads to recruitment of syntaxin 13 and strongly couples Rab4 and Rab11 domains, causing accumulation of internalized receptors in recycling endosomes. Consistent with the observed decrease in AMPAR clusters , Caspase-3 cleavage of GRASP-1 might separate the N-terminal Rab4 domain from the C-terminal syntaxin 13 binding site and disrupt the coupling between Rab4 and Rab11 domains.

    Journal: PLoS Biology

    Article Title: Neuron Specific Rab4 Effector GRASP-1 Coordinates Membrane Specialization and Maturation of Recycling Endosomes

    doi: 10.1371/journal.pbio.1000283

    Figure Lengend Snippet: Endosomes can be viewed as mosaic distribution of Rab4, Rab5, and Rab11 domains that dynamically interact via effector proteins and SNAREs. The Rab5 domain allows entry into the early/sorting endosome, whereas the Rab4 and Rab11 domains contain the machinery that is necessary for sorting and recycling membranes and receptors back to the plasma membrane. (A) GRASP-1 binds to Rab4 and syntaxin 13 and couples Rab4 and Rab11 recycling endosomes. The complex formed between GRASP-1 and t-SNARE syntaxin 13 might mediate fusion between Rab4 and Rab11 endosomes. (B) Absence of GRASP-1 interferes with complex formation at the recycling step, causing cargo accumulation in early endosomes, impairment of receptor expression, and changes in spine morphology. (C) Overexpression of GRASP-1 leads to recruitment of syntaxin 13 and strongly couples Rab4 and Rab11 domains, causing accumulation of internalized receptors in recycling endosomes. Consistent with the observed decrease in AMPAR clusters , Caspase-3 cleavage of GRASP-1 might separate the N-terminal Rab4 domain from the C-terminal syntaxin 13 binding site and disrupt the coupling between Rab4 and Rab11 domains.

    Article Snippet: The following antibodies were obtained from commercial sources: rabbit anti-GRASP-1 (AB96361), mouse anti-β-actin, mouse anti-GluR2 (Chemicon), mouse anti-Rab4, mouse anti-EEA1 (BD Biosciences), mouse anti-FLAG, mouse anti-MAP2, mouse anti-αtubulin (Sigma), mouse anti-GFP (Roche), mouse anti-bassoon (Stressgen), rabbit anti-β-galactosidase (MP Biomedicals), mouse anti-β-galactosidase (Promega), rabbit anti-GluR1 (Calbiochem), mouse anti-HA (Roche), rabbit anti-myc (Upstate Biotechnology), mouse LAMP-1 (Stressgen), mouse anti-myc, rabbit anti-Rab5 (Santa Cruz Biotechnology), rabbit anti-syntaxin 13 (Synaptic Systems), human anti-EEA1, and mouse anti-human TfR (ATCC).

    Techniques: Expressing, Over Expression, Binding Assay