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paav-hsyn-nls-cre-2a-egfp  (Addgene inc)


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    Addgene inc paav-hsyn-nls-cre-2a-egfp
    Paav Hsyn Nls Cre 2a Egfp, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. <t>AAV5-CMV-Cre-eGFP</t> was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.
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    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. <t>AAV5-CMV-Cre-eGFP</t> was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.
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    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. <t>AAV5-CMV-Cre-eGFP</t> was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.
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    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. <t>AAV5-CMV-Cre-eGFP</t> was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.
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    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. <t>AAV5-CMV-Cre-eGFP</t> was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.
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    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. <t>AAV5-CMV-Cre-eGFP</t> was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.
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    Image Search Results


    A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. AAV5-CMV-Cre-eGFP was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.

    Journal: bioRxiv

    Article Title: Deficiency of Shank3 in the Nucleus Accumbens Reveals a Loss of Social-Specific Motivation

    doi: 10.1101/2025.01.08.631742

    Figure Lengend Snippet: A) Schematic design of Shank3 deletion strategy using CRE -loxP . Arrows indicate loxP sites. Exons removed are shown in purple. AAV5-CMV-Cre-eGFP was injected directly into the NAc of Shank3 fl/fl animals to delete Δe4-22 of Shank3 . B) PCR revealed the detection of Shank3 deletion e4-22 in the NAc but not the tail in Shank3 fl/fl mice injected with Cre and Shank3 Δe – controls. C) SHANK3 protein levels were quantified using western blot analysis in eGFP or Cre-injected Shank3 fl/fl mice and Shank3 Δe – mice. SHANK3 is significantly decreased in Cre injected Shank3 fl/fl mice ( *P=0.0123 ) and Shank3 Δe – mice ( ***P=0.0008) compared to eGFP controls (Ordinary one-way ANOVA, F (2,12) =14.25; P=0.0007; n1=6, n2=6, n3=3 ). D-F ) Immunostaining of SHANK3 (red) and DAPI (blue) with eGFP (green) from viral injection. Representative 20x and 63x images of the NAc with SHANK3 immunostaining showed D ) SHANK3 was absent in Shank3 Δe – mice and E ) reduced in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP mice. F ) A secondary antibody fluorescent signal was used to visualize the SHANK3 primary antibody. Fluorescence was decreased significantly in Shank3 fl/fl+CRE mice compared to Shank3 fl/fl+eGFP (** P= 0.0078 ) and in Shank3 Δe – mice compared to Shank3 fl/fl+eGFP (** P=0.0018 ) (Ordinary one-way ANOVA, F (2,15) =11.28; P=0.0010; n1=8, n2=8, n3=6 ). Complete statistical analyses are provided in the statistical analyses file associated with this manuscript.

    Article Snippet: Shank3 fl/fl mice were injected with 300nL pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 or 300nL AAV5.hSyn.eGFP.WPRE.bGH (Addgene). pAAV.CMV.HI.eGFP-Cre.WPRE.SV40 (Addgene viral prep # 105545-AAV5; http://n2t.net/addgene:105545 ; RRID:Addgene_105545) and pAAV.CMV.PI.EGFP.WPRE.bGH (Addgene viral prep # 105530-AAV5; http://n2t.net/addgene:105530 ; RRID:Addgene_105530).

    Techniques: Injection, Western Blot, Immunostaining, Fluorescence