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Image Search Results


Striata from WT, spinophilin KO, or SAPAP3 KO mice were homogenized in RIPA (1A, 1B) or low-ionic (1B, 1C) buffer and immunoprecipitated with goat or rabbit IgG, goat spinophilin, or SAPAP3 rabbit antibody as detailed. Immunoprecipitates were immunoblotted with the same antibodies.

Journal: Molecular and cellular neurosciences

Article Title: The association of spinophilin with disks large-associated protein 3 (SAPAP3) is regulated by metabotropic glutamate receptor (mGluR) 5.

doi: 10.1016/j.mcn.2018.06.001

Figure Lengend Snippet: Striata from WT, spinophilin KO, or SAPAP3 KO mice were homogenized in RIPA (1A, 1B) or low-ionic (1B, 1C) buffer and immunoprecipitated with goat or rabbit IgG, goat spinophilin, or SAPAP3 rabbit antibody as detailed. Immunoprecipitates were immunoblotted with the same antibodies.

Article Snippet: The templates for the constructs used were: human spinophilin ( Hiday et al., 2017 ); rat SAPAP3 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P97838","term_id":"71153510","term_text":"P97838"}} P97838 ; Addgene 40217, Cambridge, MA); mouse mGluR5 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"Q3UVX5","term_id":"158705917","term_text":"Q3UVX5"}} Q3UVX5 ; Transomic Technology, Huntsville, AL {"type":"entrez-nucleotide","attrs":{"text":"BC144727","term_id":"219519512","term_text":"BC144727"}} BC144727 ); human ΡKCβ (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P05771","term_id":"20141488","term_text":"P05771"}} P05771 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC036472","term_id":"22209071","term_text":"BC036472"}} BC036472 ); human PP1α (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P62136","term_id":"49065811","term_text":"P62136"}} P62136 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC001888","term_id":"12804878","term_text":"BC001888"}} BC001888 ); and rat PP1γ1 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P63088","term_id":"52001084","term_text":"P63088"}} P63088 ; ( Carmody et al., 2008 )).

Techniques: Immunoprecipitation

HEK293 cells were transfected with HA-tagged spinophilin, V5-tagged SAPAP3, and Myc-tagged mGluR5. A. Transfected lysates were immunoprecipitated with HA or Myc antibodies and immunoblotted with spinophilin, SAPAP3, and mGluR5 antibodies. B. Overexpression of mGluR5 significantly increased the amount of SAPAP3 in spinophilin immunoprecipitates. C. Overexpression of spinophilin had no significant effect on the association of SAPAP3 with mGluR5. D. SAPAP3 overexpression significantly enhanced the association of spinophilin with mGluR5. E. Schema of SAPAP3/mGluR5-dependent bridging of spinophilin. ** p<0.01, *** p <0.001.

Journal: Molecular and cellular neurosciences

Article Title: The association of spinophilin with disks large-associated protein 3 (SAPAP3) is regulated by metabotropic glutamate receptor (mGluR) 5.

doi: 10.1016/j.mcn.2018.06.001

Figure Lengend Snippet: HEK293 cells were transfected with HA-tagged spinophilin, V5-tagged SAPAP3, and Myc-tagged mGluR5. A. Transfected lysates were immunoprecipitated with HA or Myc antibodies and immunoblotted with spinophilin, SAPAP3, and mGluR5 antibodies. B. Overexpression of mGluR5 significantly increased the amount of SAPAP3 in spinophilin immunoprecipitates. C. Overexpression of spinophilin had no significant effect on the association of SAPAP3 with mGluR5. D. SAPAP3 overexpression significantly enhanced the association of spinophilin with mGluR5. E. Schema of SAPAP3/mGluR5-dependent bridging of spinophilin. ** p<0.01, *** p <0.001.

Article Snippet: The templates for the constructs used were: human spinophilin ( Hiday et al., 2017 ); rat SAPAP3 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P97838","term_id":"71153510","term_text":"P97838"}} P97838 ; Addgene 40217, Cambridge, MA); mouse mGluR5 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"Q3UVX5","term_id":"158705917","term_text":"Q3UVX5"}} Q3UVX5 ; Transomic Technology, Huntsville, AL {"type":"entrez-nucleotide","attrs":{"text":"BC144727","term_id":"219519512","term_text":"BC144727"}} BC144727 ); human ΡKCβ (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P05771","term_id":"20141488","term_text":"P05771"}} P05771 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC036472","term_id":"22209071","term_text":"BC036472"}} BC036472 ); human PP1α (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P62136","term_id":"49065811","term_text":"P62136"}} P62136 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC001888","term_id":"12804878","term_text":"BC001888"}} BC001888 ); and rat PP1γ1 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P63088","term_id":"52001084","term_text":"P63088"}} P63088 ; ( Carmody et al., 2008 )).

Techniques: Transfection, Immunoprecipitation, Over Expression

HEK293 cells were transfected with SAPAP3 and mGluR5 along with different deletion mutants of spinophilin. A. Schematic representation of spinophilin deletion mutants. Numbers refer to amino acids on different spinophilin fragments. B. HEK293 cells were transfected with SAPAP3 and mGluR5 alone or in the presence of different HA-tagged spinophilin deletion mutants. HEK293 cell lysates were immunoprecipitated with an HA antibody and immunoprecipitates were immunoblotted for SAPAP3, mGluR5, and HA. C. A different set of HA spinophilin fragments were immunoprecipitated and immunoblotted for HA and mGluR5. Experiments are representative of 3 independent trials.

Journal: Molecular and cellular neurosciences

Article Title: The association of spinophilin with disks large-associated protein 3 (SAPAP3) is regulated by metabotropic glutamate receptor (mGluR) 5.

doi: 10.1016/j.mcn.2018.06.001

Figure Lengend Snippet: HEK293 cells were transfected with SAPAP3 and mGluR5 along with different deletion mutants of spinophilin. A. Schematic representation of spinophilin deletion mutants. Numbers refer to amino acids on different spinophilin fragments. B. HEK293 cells were transfected with SAPAP3 and mGluR5 alone or in the presence of different HA-tagged spinophilin deletion mutants. HEK293 cell lysates were immunoprecipitated with an HA antibody and immunoprecipitates were immunoblotted for SAPAP3, mGluR5, and HA. C. A different set of HA spinophilin fragments were immunoprecipitated and immunoblotted for HA and mGluR5. Experiments are representative of 3 independent trials.

Article Snippet: The templates for the constructs used were: human spinophilin ( Hiday et al., 2017 ); rat SAPAP3 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P97838","term_id":"71153510","term_text":"P97838"}} P97838 ; Addgene 40217, Cambridge, MA); mouse mGluR5 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"Q3UVX5","term_id":"158705917","term_text":"Q3UVX5"}} Q3UVX5 ; Transomic Technology, Huntsville, AL {"type":"entrez-nucleotide","attrs":{"text":"BC144727","term_id":"219519512","term_text":"BC144727"}} BC144727 ); human ΡKCβ (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P05771","term_id":"20141488","term_text":"P05771"}} P05771 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC036472","term_id":"22209071","term_text":"BC036472"}} BC036472 ); human PP1α (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P62136","term_id":"49065811","term_text":"P62136"}} P62136 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC001888","term_id":"12804878","term_text":"BC001888"}} BC001888 ); and rat PP1γ1 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P63088","term_id":"52001084","term_text":"P63088"}} P63088 ; ( Carmody et al., 2008 )).

Techniques: Transfection, Immunoprecipitation

HEK293 cells were transfected with spinophilin and SAPAP3 in the absence or presence of PKCβ. The following day, HEK293 cells were treated with 200 nM TPA or vehicle for 30 minutes. A. Lysates were immunoprecipitated with an HA spinophilin antibody and lysates and immunoprecipitates were stained with Ponceau S and immunoblotted for spinophilin, SAPAP3, PKCβ, and PKC substrate substrate antibody. B. Activation of PKC and overexpression of PKCβ along with activation of PKC significantly decreased the expression of SAPAP3 compared to: no overexpression or activation (* symbol), PKCβ overexpression only (# symbol), or activation only ($ symbol). C. Activation of PKC along with overexpression of PKCβ significantly increased the association between spinophilin and SAPAP3. *,#,$ p<0.05, $$ p<0.01, ***, ###p <0.001, **** p<0.0001.

Journal: Molecular and cellular neurosciences

Article Title: The association of spinophilin with disks large-associated protein 3 (SAPAP3) is regulated by metabotropic glutamate receptor (mGluR) 5.

doi: 10.1016/j.mcn.2018.06.001

Figure Lengend Snippet: HEK293 cells were transfected with spinophilin and SAPAP3 in the absence or presence of PKCβ. The following day, HEK293 cells were treated with 200 nM TPA or vehicle for 30 minutes. A. Lysates were immunoprecipitated with an HA spinophilin antibody and lysates and immunoprecipitates were stained with Ponceau S and immunoblotted for spinophilin, SAPAP3, PKCβ, and PKC substrate substrate antibody. B. Activation of PKC and overexpression of PKCβ along with activation of PKC significantly decreased the expression of SAPAP3 compared to: no overexpression or activation (* symbol), PKCβ overexpression only (# symbol), or activation only ($ symbol). C. Activation of PKC along with overexpression of PKCβ significantly increased the association between spinophilin and SAPAP3. *,#,$ p<0.05, $$ p<0.01, ***, ###p <0.001, **** p<0.0001.

Article Snippet: The templates for the constructs used were: human spinophilin ( Hiday et al., 2017 ); rat SAPAP3 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P97838","term_id":"71153510","term_text":"P97838"}} P97838 ; Addgene 40217, Cambridge, MA); mouse mGluR5 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"Q3UVX5","term_id":"158705917","term_text":"Q3UVX5"}} Q3UVX5 ; Transomic Technology, Huntsville, AL {"type":"entrez-nucleotide","attrs":{"text":"BC144727","term_id":"219519512","term_text":"BC144727"}} BC144727 ); human ΡKCβ (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P05771","term_id":"20141488","term_text":"P05771"}} P05771 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC036472","term_id":"22209071","term_text":"BC036472"}} BC036472 ); human PP1α (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P62136","term_id":"49065811","term_text":"P62136"}} P62136 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC001888","term_id":"12804878","term_text":"BC001888"}} BC001888 ); and rat PP1γ1 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P63088","term_id":"52001084","term_text":"P63088"}} P63088 ; ( Carmody et al., 2008 )).

Techniques: Transfection, Immunoprecipitation, Staining, Activation Assay, Over Expression, Expressing

HEK293 cells were transfected with SAPAP3 and mGluR5 and/or spinophilin in the absence or presence of PPlα. A. HA immunoprecipitates and/or lysates were immunoblotted with spinophilin, SAPAP3 and Myc antibodies. B. Overexpression of PP1 significantly decreased SAPAP3 association with spinophilin. C. HA immunoprecipitates and/or lysates were immunoblotted with spinophilin, mGluR5, and Myc antibodies. D. Overexpression of PP1 decreased mGluR association with spinophilin. E. WT or F451A (FA) mutant spinophilin were transfected with SAPAP3 and mGluR5. Mutant spinophilin did not bind endogenous PP1, but had no effect on the association of spinophilin with SAPAP3 (F) or mGluR5 (G). **P<0.01, ****P<0.0001.

Journal: Molecular and cellular neurosciences

Article Title: The association of spinophilin with disks large-associated protein 3 (SAPAP3) is regulated by metabotropic glutamate receptor (mGluR) 5.

doi: 10.1016/j.mcn.2018.06.001

Figure Lengend Snippet: HEK293 cells were transfected with SAPAP3 and mGluR5 and/or spinophilin in the absence or presence of PPlα. A. HA immunoprecipitates and/or lysates were immunoblotted with spinophilin, SAPAP3 and Myc antibodies. B. Overexpression of PP1 significantly decreased SAPAP3 association with spinophilin. C. HA immunoprecipitates and/or lysates were immunoblotted with spinophilin, mGluR5, and Myc antibodies. D. Overexpression of PP1 decreased mGluR association with spinophilin. E. WT or F451A (FA) mutant spinophilin were transfected with SAPAP3 and mGluR5. Mutant spinophilin did not bind endogenous PP1, but had no effect on the association of spinophilin with SAPAP3 (F) or mGluR5 (G). **P<0.01, ****P<0.0001.

Article Snippet: The templates for the constructs used were: human spinophilin ( Hiday et al., 2017 ); rat SAPAP3 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P97838","term_id":"71153510","term_text":"P97838"}} P97838 ; Addgene 40217, Cambridge, MA); mouse mGluR5 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"Q3UVX5","term_id":"158705917","term_text":"Q3UVX5"}} Q3UVX5 ; Transomic Technology, Huntsville, AL {"type":"entrez-nucleotide","attrs":{"text":"BC144727","term_id":"219519512","term_text":"BC144727"}} BC144727 ); human ΡKCβ (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P05771","term_id":"20141488","term_text":"P05771"}} P05771 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC036472","term_id":"22209071","term_text":"BC036472"}} BC036472 ); human PP1α (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P62136","term_id":"49065811","term_text":"P62136"}} P62136 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC001888","term_id":"12804878","term_text":"BC001888"}} BC001888 ); and rat PP1γ1 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P63088","term_id":"52001084","term_text":"P63088"}} P63088 ; ( Carmody et al., 2008 )).

Techniques: Transfection, Over Expression, Mutagenesis

Striatal slices were incubated with DHPG or vehicle for 30 minutes. Following incubation, striata were dissected and immunoprecipitated for spinophilin and SAPAP3. A. Immunoblots of control and treated striatal lysates and spinophilin and SAPAP3 immunoprecipitates. B. Activation of group I mGluRs led to a trend towards an increase in the association of spinophilin with SAPAP3.

Journal: Molecular and cellular neurosciences

Article Title: The association of spinophilin with disks large-associated protein 3 (SAPAP3) is regulated by metabotropic glutamate receptor (mGluR) 5.

doi: 10.1016/j.mcn.2018.06.001

Figure Lengend Snippet: Striatal slices were incubated with DHPG or vehicle for 30 minutes. Following incubation, striata were dissected and immunoprecipitated for spinophilin and SAPAP3. A. Immunoblots of control and treated striatal lysates and spinophilin and SAPAP3 immunoprecipitates. B. Activation of group I mGluRs led to a trend towards an increase in the association of spinophilin with SAPAP3.

Article Snippet: The templates for the constructs used were: human spinophilin ( Hiday et al., 2017 ); rat SAPAP3 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P97838","term_id":"71153510","term_text":"P97838"}} P97838 ; Addgene 40217, Cambridge, MA); mouse mGluR5 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"Q3UVX5","term_id":"158705917","term_text":"Q3UVX5"}} Q3UVX5 ; Transomic Technology, Huntsville, AL {"type":"entrez-nucleotide","attrs":{"text":"BC144727","term_id":"219519512","term_text":"BC144727"}} BC144727 ); human ΡKCβ (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P05771","term_id":"20141488","term_text":"P05771"}} P05771 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC036472","term_id":"22209071","term_text":"BC036472"}} BC036472 ); human PP1α (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P62136","term_id":"49065811","term_text":"P62136"}} P62136 ; Transomic Technology {"type":"entrez-nucleotide","attrs":{"text":"BC001888","term_id":"12804878","term_text":"BC001888"}} BC001888 ); and rat PP1γ1 (Uniprot ID {"type":"entrez-protein","attrs":{"text":"P63088","term_id":"52001084","term_text":"P63088"}} P63088 ; ( Carmody et al., 2008 )).

Techniques: Incubation, Immunoprecipitation, Western Blot, Activation Assay