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gfp sbp rtn4a  (Addgene inc)

 
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    Addgene inc gfp sbp rtn4a
    Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or <t>V5-AP-RTN4A</t> as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also <xref ref-type=Figure S3 and Table S3 . " width="250" height="auto" />
    Gfp Sbp Rtn4a, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Axonal endoplasmic reticulum tubules control local translation via P180/RRBP1-mediated ribosome interactions"

    Article Title: Axonal endoplasmic reticulum tubules control local translation via P180/RRBP1-mediated ribosome interactions

    Journal: Developmental Cell

    doi: 10.1016/j.devcel.2024.05.005

    Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or V5-AP-RTN4A as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also <xref ref-type=Figure S3 and Table S3 . " title="... from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or V5-AP-RTN4A as a negative control (right). Expression of constructs ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or V5-AP-RTN4A as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also Figure S3 and Table S3 .

    Techniques Used: Expressing, Negative Control, Construct, MANN-WHITNEY, Control


    Figure Legend Snippet:

    Techniques Used: Recombinant, Magnetic Beads, shRNA, Sequencing, Software



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    Addgene inc gfp sbp rtn4a
    Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or <t>V5-AP-RTN4A</t> as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also <xref ref-type=Figure S3 and Table S3 . " width="250" height="auto" />
    Gfp Sbp Rtn4a, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gfp sbp rtn4a/product/Addgene inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    gfp sbp rtn4a - by Bioz Stars, 2024-10
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    n a gfp sbp rtn4a farı́as  (Addgene inc)
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    Addgene inc n a gfp sbp rtn4a farı́as
    Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or <t>V5-AP-RTN4A</t> as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also <xref ref-type=Figure S3 and Table S3 . " width="250" height="auto" />
    N A Gfp Sbp Rtn4a Farı́as, supplied by Addgene inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/n a gfp sbp rtn4a farı́as/product/Addgene inc
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    n a gfp sbp rtn4a farı́as - by Bioz Stars, 2024-10
    86/100 stars
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    Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or V5-AP-RTN4A as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also <xref ref-type=Figure S3 and Table S3 . " width="100%" height="100%">

    Journal: Developmental Cell

    Article Title: Axonal endoplasmic reticulum tubules control local translation via P180/RRBP1-mediated ribosome interactions

    doi: 10.1016/j.devcel.2024.05.005

    Figure Lengend Snippet: Axonal ER-ribosome interactions are enriched at axon branch points and regulated by extrinsic cues (A–C) Schematic representation of split APEX system used to detect ER-ribosome contacts. When the ER protein Sec61β fused to AP module and the ribosomal protein RpL10A fused to EX module interact with each other, APEX is reconstituted, and contact sites can be visualized as a biotinylation radius around the interactions (A). Representative images of split APEX assay in distal axons from neurons expressing RpL10A-3xHA-EX and V5-AP-Sec61β (left) or V5-AP-RTN4A as a negative control (right). Expression of constructs is visualized with V5 and HA antibodies, and biotinylation is detected with conjugated Strep-555 (B). Quantification of Strep signal in distal axons from neurons as in (B) and without H2O2 as a negative control for the biotinylation reaction (C). (D) Representative image of an axon with branches of a DIV7 neuron expressing V5-AP-Sec61β and RpL10A-3xHA-EX DIV7, showing enrichment of split APEX signal at branch points. White arrowheads indicate branch points. Split APEX signal (Strep555) is shown in magenta and a high-low intensity scale as indicated in lower panel. (E) Quantification of axonal ER-bound ribosomes, using split APEX assay as in (D), analyzed in the axon shaft and at axonal branch points. n = 16 per condition re-analyzed from six independent experiments. ∗ p < 0.05 comparing conditions to each other using Mann-Whitney tests. Scale bars represent 5 μm. (F) Quantification of axonal ER-bound ribosomes, using split APEX assay with RpL10A-3xHA-EX and V5-AP-Sec61β, in neurons stimulated for 30 min with BSA (control), BDNF, NT-3, or NGF. Individual data points each represent a neuron in (C), (E), and (F), and each color represents an independent experiment in (F). Data are presented as mean values ± SEM in (C), (E), and (F). ns, not significant; ∗ p < 0.05; ∗∗ p < 0.01; and ∗∗∗ p < 0.001 comparing conditions to control using unpaired t tests (C), Mann-Whitney test (E), or ordinary one-way ANOVA tests (F). Scale bars represent 5 μm (B) and (D). See also Figure S3 and Table S3 .

    Article Snippet: Following vectors were used: pSuper, pGW1-mCherry and pGW1-BFP, RTN4A-GFP (a gift from Dr. Gia Voeltz, Addgene plasmid #61807), pEGFP(A206K)-N1 and pEGFP(A206K)-C1 (a gift from Dr. Jennifer Lippincott-Schwartz), GFP-Sec61β (a gift from Dr. Tom Rapoport, Addgene # 15108), RpL10A-tagRFP (a gift from dr. Robert Singer, Addgene #74172), TOM20-V5-FKBP-AP (a gift from Alice Ting, Addgene#120914), mCherry myr 5’/3’-Calreticulin (a gift from dr. Jefferey L. Twiss), Lifeact-mCherry (a gift from dr. Moritoshi Sato, Addgene #67302), HA-KifC1-MD-Strep, DP1-GFP, GFP-SBP-RTN4A, P180-ΔCoiled-coil, P180-Δrepeats-GFP, P180-Coiled-coil and P180-Repeats were previously described.

    Techniques: Expressing, Negative Control, Construct, MANN-WHITNEY, Control

    Journal: Developmental Cell

    Article Title: Axonal endoplasmic reticulum tubules control local translation via P180/RRBP1-mediated ribosome interactions

    doi: 10.1016/j.devcel.2024.05.005

    Figure Lengend Snippet:

    Article Snippet: Following vectors were used: pSuper, pGW1-mCherry and pGW1-BFP, RTN4A-GFP (a gift from Dr. Gia Voeltz, Addgene plasmid #61807), pEGFP(A206K)-N1 and pEGFP(A206K)-C1 (a gift from Dr. Jennifer Lippincott-Schwartz), GFP-Sec61β (a gift from Dr. Tom Rapoport, Addgene # 15108), RpL10A-tagRFP (a gift from dr. Robert Singer, Addgene #74172), TOM20-V5-FKBP-AP (a gift from Alice Ting, Addgene#120914), mCherry myr 5’/3’-Calreticulin (a gift from dr. Jefferey L. Twiss), Lifeact-mCherry (a gift from dr. Moritoshi Sato, Addgene #67302), HA-KifC1-MD-Strep, DP1-GFP, GFP-SBP-RTN4A, P180-ΔCoiled-coil, P180-Δrepeats-GFP, P180-Coiled-coil and P180-Repeats were previously described.

    Techniques: Recombinant, Magnetic Beads, shRNA, Sequencing, Software