pcmv6 cacna1b myc ddk  (OriGene)


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    Structured Review

    OriGene pcmv6 cacna1b myc ddk
    <t>Cav2.2</t> protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01
    Pcmv6 Cacna1b Myc Ddk, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcmv6 cacna1b myc ddk/product/OriGene
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcmv6 cacna1b myc ddk - by Bioz Stars, 2024-06
    91/100 stars

    Images

    1) Product Images from "Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain"

    Article Title: Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain

    Journal: Molecular Neurobiology

    doi: 10.1007/s12035-020-02276-8

    Cav2.2 protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01
    Figure Legend Snippet: Cav2.2 protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01

    Techniques Used: Expressing

    Sigma-1 receptor transcription, protein expression, and surface level do not change after SNI in rat DRGs. a Sample blots of sigma-1 receptor (Sig-1R) expression at different time points after SNI. α-Tubulin served as internal control. b Summary data show that Sig-1R protein expression decreases slightly at day 14 (nonsignificant per ANOVA) but not at days 3 or 28 after SNI (left panel). The Sig-1R mRNA does not change after SNI at days 3, 14, or 28 (right panel). c Surface level of Sig-1Rs remains unchanged after SNI. Streptavidin beads were used to pull down biotinylated lysate (200 μg) from DRGs 14 days after SNI or sham surgery. Na + -K + ATPase served as internal control and GAPDH served as negative control. d Sig-1R KO increases Cav2.2 expression in HEK cells. Western blotting shows an increase of Cav2.2 protein level in total cellular lysates of Sig-1R KO HEK cells when compared to wild-type cells. Data represent means ± SEM; two-way ANOVA or Student’s t test
    Figure Legend Snippet: Sigma-1 receptor transcription, protein expression, and surface level do not change after SNI in rat DRGs. a Sample blots of sigma-1 receptor (Sig-1R) expression at different time points after SNI. α-Tubulin served as internal control. b Summary data show that Sig-1R protein expression decreases slightly at day 14 (nonsignificant per ANOVA) but not at days 3 or 28 after SNI (left panel). The Sig-1R mRNA does not change after SNI at days 3, 14, or 28 (right panel). c Surface level of Sig-1Rs remains unchanged after SNI. Streptavidin beads were used to pull down biotinylated lysate (200 μg) from DRGs 14 days after SNI or sham surgery. Na + -K + ATPase served as internal control and GAPDH served as negative control. d Sig-1R KO increases Cav2.2 expression in HEK cells. Western blotting shows an increase of Cav2.2 protein level in total cellular lysates of Sig-1R KO HEK cells when compared to wild-type cells. Data represent means ± SEM; two-way ANOVA or Student’s t test

    Techniques Used: Expressing, Negative Control, Western Blot

    cFOS increases 4E-BP1 transcription via its interaction with Sig-1R. a Overexpression of 4E-BP1 decreases Cav2.2 in HEK cells. b Sig-1R interacts with cFOS in HEK cells. In cFOS-Myc/DDK and Sig-1R-GFP HEK co-expressing HEK cells, sample blot demonstrates the co-immunoprecipitation of those two proteins (lane 6). c Co-overexpressed Sig-1R and cFOS bind to the promoter of 4E-BP1. DNAs obtained from chromatin immunoprecipitation assays testing the binding of GFP-tagged Sig-1R (upper panel) or Myc-tagged cFOS (lower panel) to the 4E-BP1 promoter in HEK cells were examined. The precipitated DNA was amplified using specific primers for the 4E-BP1 promoter. Quantified results from the PCR products were normalized to input. Summary data are presented as means ± SEM; Student’s t test; * P < 0.05, *** P < 0.001; N = 3
    Figure Legend Snippet: cFOS increases 4E-BP1 transcription via its interaction with Sig-1R. a Overexpression of 4E-BP1 decreases Cav2.2 in HEK cells. b Sig-1R interacts with cFOS in HEK cells. In cFOS-Myc/DDK and Sig-1R-GFP HEK co-expressing HEK cells, sample blot demonstrates the co-immunoprecipitation of those two proteins (lane 6). c Co-overexpressed Sig-1R and cFOS bind to the promoter of 4E-BP1. DNAs obtained from chromatin immunoprecipitation assays testing the binding of GFP-tagged Sig-1R (upper panel) or Myc-tagged cFOS (lower panel) to the 4E-BP1 promoter in HEK cells were examined. The precipitated DNA was amplified using specific primers for the 4E-BP1 promoter. Quantified results from the PCR products were normalized to input. Summary data are presented as means ± SEM; Student’s t test; * P < 0.05, *** P < 0.001; N = 3

    Techniques Used: Over Expression, Expressing, Immunoprecipitation, Chromatin Immunoprecipitation, Binding Assay, Amplification

    Sig-1R regulates 4E-BP1 and its effect on eIF4E. a Sig-1R KO decreases the transcription and translation of 4E-BP1 in HEK cells. Sample blot demonstrates a decrease of 4E-BP1 protein in Sig-1R KO HEK cells. Summary data reveal an elevated level of protein and mRNA of 4E-BP1 in Sig-1R KO HEK cells. b Sig-1R knockout (KO) increases eIF4E and Cav2.2 mRNA binding. In the RNA-IP test, eIF4E binding to Cav2.2 5′cap region increases in Sig-1R KO HEK cells. Data represent means ± SEM; Student’s t test; Mann-Whitney test for b ; * P < 0.05, *** P < 0.001
    Figure Legend Snippet: Sig-1R regulates 4E-BP1 and its effect on eIF4E. a Sig-1R KO decreases the transcription and translation of 4E-BP1 in HEK cells. Sample blot demonstrates a decrease of 4E-BP1 protein in Sig-1R KO HEK cells. Summary data reveal an elevated level of protein and mRNA of 4E-BP1 in Sig-1R KO HEK cells. b Sig-1R knockout (KO) increases eIF4E and Cav2.2 mRNA binding. In the RNA-IP test, eIF4E binding to Cav2.2 5′cap region increases in Sig-1R KO HEK cells. Data represent means ± SEM; Student’s t test; Mann-Whitney test for b ; * P < 0.05, *** P < 0.001

    Techniques Used: Knock-Out, Binding Assay, MANN-WHITNEY

    pcmv6 cacna1b myc ddk  (OriGene)


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    Structured Review

    OriGene pcmv6 cacna1b myc ddk
    Pcmv6 Cacna1b Myc Ddk, supplied by OriGene, 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/pcmv6 cacna1b myc ddk/product/OriGene
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcmv6 cacna1b myc ddk - by Bioz Stars, 2024-06
    86/100 stars

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    pcmv6 cacna1b myc ddk  (OriGene)


    Bioz Verified Symbol OriGene is a verified supplier
    Bioz Manufacturer Symbol OriGene manufactures this product  
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  • 91

    Structured Review

    OriGene pcmv6 cacna1b myc ddk
    <t>Cav2.2</t> protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01
    Pcmv6 Cacna1b Myc Ddk, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcmv6 cacna1b myc ddk/product/OriGene
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcmv6 cacna1b myc ddk - by Bioz Stars, 2024-06
    91/100 stars

    Images

    1) Product Images from "Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain"

    Article Title: Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain

    Journal: Molecular Neurobiology

    doi: 10.1007/s12035-020-02276-8

    Cav2.2 protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01
    Figure Legend Snippet: Cav2.2 protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01

    Techniques Used: Expressing

    Sigma-1 receptor transcription, protein expression, and surface level do not change after SNI in rat DRGs. a Sample blots of sigma-1 receptor (Sig-1R) expression at different time points after SNI. α-Tubulin served as internal control. b Summary data show that Sig-1R protein expression decreases slightly at day 14 (nonsignificant per ANOVA) but not at days 3 or 28 after SNI (left panel). The Sig-1R mRNA does not change after SNI at days 3, 14, or 28 (right panel). c Surface level of Sig-1Rs remains unchanged after SNI. Streptavidin beads were used to pull down biotinylated lysate (200 μg) from DRGs 14 days after SNI or sham surgery. Na + -K + ATPase served as internal control and GAPDH served as negative control. d Sig-1R KO increases Cav2.2 expression in HEK cells. Western blotting shows an increase of Cav2.2 protein level in total cellular lysates of Sig-1R KO HEK cells when compared to wild-type cells. Data represent means ± SEM; two-way ANOVA or Student’s t test
    Figure Legend Snippet: Sigma-1 receptor transcription, protein expression, and surface level do not change after SNI in rat DRGs. a Sample blots of sigma-1 receptor (Sig-1R) expression at different time points after SNI. α-Tubulin served as internal control. b Summary data show that Sig-1R protein expression decreases slightly at day 14 (nonsignificant per ANOVA) but not at days 3 or 28 after SNI (left panel). The Sig-1R mRNA does not change after SNI at days 3, 14, or 28 (right panel). c Surface level of Sig-1Rs remains unchanged after SNI. Streptavidin beads were used to pull down biotinylated lysate (200 μg) from DRGs 14 days after SNI or sham surgery. Na + -K + ATPase served as internal control and GAPDH served as negative control. d Sig-1R KO increases Cav2.2 expression in HEK cells. Western blotting shows an increase of Cav2.2 protein level in total cellular lysates of Sig-1R KO HEK cells when compared to wild-type cells. Data represent means ± SEM; two-way ANOVA or Student’s t test

    Techniques Used: Expressing, Negative Control, Western Blot

    cFOS increases 4E-BP1 transcription via its interaction with Sig-1R. a Overexpression of 4E-BP1 decreases Cav2.2 in HEK cells. b Sig-1R interacts with cFOS in HEK cells. In cFOS-Myc/DDK and Sig-1R-GFP HEK co-expressing HEK cells, sample blot demonstrates the co-immunoprecipitation of those two proteins (lane 6). c Co-overexpressed Sig-1R and cFOS bind to the promoter of 4E-BP1. DNAs obtained from chromatin immunoprecipitation assays testing the binding of GFP-tagged Sig-1R (upper panel) or Myc-tagged cFOS (lower panel) to the 4E-BP1 promoter in HEK cells were examined. The precipitated DNA was amplified using specific primers for the 4E-BP1 promoter. Quantified results from the PCR products were normalized to input. Summary data are presented as means ± SEM; Student’s t test; * P < 0.05, *** P < 0.001; N = 3
    Figure Legend Snippet: cFOS increases 4E-BP1 transcription via its interaction with Sig-1R. a Overexpression of 4E-BP1 decreases Cav2.2 in HEK cells. b Sig-1R interacts with cFOS in HEK cells. In cFOS-Myc/DDK and Sig-1R-GFP HEK co-expressing HEK cells, sample blot demonstrates the co-immunoprecipitation of those two proteins (lane 6). c Co-overexpressed Sig-1R and cFOS bind to the promoter of 4E-BP1. DNAs obtained from chromatin immunoprecipitation assays testing the binding of GFP-tagged Sig-1R (upper panel) or Myc-tagged cFOS (lower panel) to the 4E-BP1 promoter in HEK cells were examined. The precipitated DNA was amplified using specific primers for the 4E-BP1 promoter. Quantified results from the PCR products were normalized to input. Summary data are presented as means ± SEM; Student’s t test; * P < 0.05, *** P < 0.001; N = 3

    Techniques Used: Over Expression, Expressing, Immunoprecipitation, Chromatin Immunoprecipitation, Binding Assay, Amplification

    Sig-1R regulates 4E-BP1 and its effect on eIF4E. a Sig-1R KO decreases the transcription and translation of 4E-BP1 in HEK cells. Sample blot demonstrates a decrease of 4E-BP1 protein in Sig-1R KO HEK cells. Summary data reveal an elevated level of protein and mRNA of 4E-BP1 in Sig-1R KO HEK cells. b Sig-1R knockout (KO) increases eIF4E and Cav2.2 mRNA binding. In the RNA-IP test, eIF4E binding to Cav2.2 5′cap region increases in Sig-1R KO HEK cells. Data represent means ± SEM; Student’s t test; Mann-Whitney test for b ; * P < 0.05, *** P < 0.001
    Figure Legend Snippet: Sig-1R regulates 4E-BP1 and its effect on eIF4E. a Sig-1R KO decreases the transcription and translation of 4E-BP1 in HEK cells. Sample blot demonstrates a decrease of 4E-BP1 protein in Sig-1R KO HEK cells. Summary data reveal an elevated level of protein and mRNA of 4E-BP1 in Sig-1R KO HEK cells. b Sig-1R knockout (KO) increases eIF4E and Cav2.2 mRNA binding. In the RNA-IP test, eIF4E binding to Cav2.2 5′cap region increases in Sig-1R KO HEK cells. Data represent means ± SEM; Student’s t test; Mann-Whitney test for b ; * P < 0.05, *** P < 0.001

    Techniques Used: Knock-Out, Binding Assay, MANN-WHITNEY

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    OriGene pcmv6 cacna1b myc ddk
    <t>Cav2.2</t> protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01
    Pcmv6 Cacna1b Myc Ddk, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcmv6 cacna1b myc ddk/product/OriGene
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcmv6 cacna1b myc ddk - by Bioz Stars, 2024-06
    91/100 stars
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    Cav2.2 protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01

    Journal: Molecular Neurobiology

    Article Title: Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain

    doi: 10.1007/s12035-020-02276-8

    Figure Lengend Snippet: Cav2.2 protein expression is downregulated in DRGs after SNI. a Total Cav2.2 protein expression deceases after SNI. Sample bands of Cav2.2 expression were demonstrated at different time after sham surgery or SNI from rat DRGs. α-Tubulin served as internal control. Summary data showed that Cav2.2 protein expression but not mRNA level decreases after SNI on day 3, 14, or 28. b Cav2.2 protein level decreases at the plasma membrane after SNI. Cav2.2 at plasma membrane decreases after day 14 of SNI. N-Cadherin expression from streptavidin beads pulled down served as internal control. Data are means ± SEM; two-way ANOVA, or Student’s t test; * P < 0.05, ** P < 0.01

    Article Snippet: Plasmid vectors used were pCMV6-CACNA1B-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC217170), pCMV6-EIF4EBP1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC201348), pCMV6-FOS-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC202597), pCMV6-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. PS100001), pCMV6-MZF1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC220791), pCMV6-OCT1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC208599), pCMV3-GFP control vector (Sino Biological, Wayne, PA; Cat. CV026), pCMV3-HA-Sec61β□ (Sino Biological, Wayne, PA; Cat. MG51753-NY), pCMV3-HA-SP3 (Sino Biological, Wayne, PA; Cat. HG13964-CY), pCMV3-Sigma-1R-GFP (Sino Biological, Wayne, PA; Cat. MG57873-ACG), pCMV3-HA control vector (Sino Biological, Wayne, PA; Cat. CV017), pcDNA3-HA-CEBPD (self-construct), pcDNA3-HA-SOX2 (self-construct), and Sigma-1R-EYFP (self-construct).

    Techniques: Expressing

    Sigma-1 receptor transcription, protein expression, and surface level do not change after SNI in rat DRGs. a Sample blots of sigma-1 receptor (Sig-1R) expression at different time points after SNI. α-Tubulin served as internal control. b Summary data show that Sig-1R protein expression decreases slightly at day 14 (nonsignificant per ANOVA) but not at days 3 or 28 after SNI (left panel). The Sig-1R mRNA does not change after SNI at days 3, 14, or 28 (right panel). c Surface level of Sig-1Rs remains unchanged after SNI. Streptavidin beads were used to pull down biotinylated lysate (200 μg) from DRGs 14 days after SNI or sham surgery. Na + -K + ATPase served as internal control and GAPDH served as negative control. d Sig-1R KO increases Cav2.2 expression in HEK cells. Western blotting shows an increase of Cav2.2 protein level in total cellular lysates of Sig-1R KO HEK cells when compared to wild-type cells. Data represent means ± SEM; two-way ANOVA or Student’s t test

    Journal: Molecular Neurobiology

    Article Title: Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain

    doi: 10.1007/s12035-020-02276-8

    Figure Lengend Snippet: Sigma-1 receptor transcription, protein expression, and surface level do not change after SNI in rat DRGs. a Sample blots of sigma-1 receptor (Sig-1R) expression at different time points after SNI. α-Tubulin served as internal control. b Summary data show that Sig-1R protein expression decreases slightly at day 14 (nonsignificant per ANOVA) but not at days 3 or 28 after SNI (left panel). The Sig-1R mRNA does not change after SNI at days 3, 14, or 28 (right panel). c Surface level of Sig-1Rs remains unchanged after SNI. Streptavidin beads were used to pull down biotinylated lysate (200 μg) from DRGs 14 days after SNI or sham surgery. Na + -K + ATPase served as internal control and GAPDH served as negative control. d Sig-1R KO increases Cav2.2 expression in HEK cells. Western blotting shows an increase of Cav2.2 protein level in total cellular lysates of Sig-1R KO HEK cells when compared to wild-type cells. Data represent means ± SEM; two-way ANOVA or Student’s t test

    Article Snippet: Plasmid vectors used were pCMV6-CACNA1B-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC217170), pCMV6-EIF4EBP1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC201348), pCMV6-FOS-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC202597), pCMV6-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. PS100001), pCMV6-MZF1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC220791), pCMV6-OCT1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC208599), pCMV3-GFP control vector (Sino Biological, Wayne, PA; Cat. CV026), pCMV3-HA-Sec61β□ (Sino Biological, Wayne, PA; Cat. MG51753-NY), pCMV3-HA-SP3 (Sino Biological, Wayne, PA; Cat. HG13964-CY), pCMV3-Sigma-1R-GFP (Sino Biological, Wayne, PA; Cat. MG57873-ACG), pCMV3-HA control vector (Sino Biological, Wayne, PA; Cat. CV017), pcDNA3-HA-CEBPD (self-construct), pcDNA3-HA-SOX2 (self-construct), and Sigma-1R-EYFP (self-construct).

    Techniques: Expressing, Negative Control, Western Blot

    cFOS increases 4E-BP1 transcription via its interaction with Sig-1R. a Overexpression of 4E-BP1 decreases Cav2.2 in HEK cells. b Sig-1R interacts with cFOS in HEK cells. In cFOS-Myc/DDK and Sig-1R-GFP HEK co-expressing HEK cells, sample blot demonstrates the co-immunoprecipitation of those two proteins (lane 6). c Co-overexpressed Sig-1R and cFOS bind to the promoter of 4E-BP1. DNAs obtained from chromatin immunoprecipitation assays testing the binding of GFP-tagged Sig-1R (upper panel) or Myc-tagged cFOS (lower panel) to the 4E-BP1 promoter in HEK cells were examined. The precipitated DNA was amplified using specific primers for the 4E-BP1 promoter. Quantified results from the PCR products were normalized to input. Summary data are presented as means ± SEM; Student’s t test; * P < 0.05, *** P < 0.001; N = 3

    Journal: Molecular Neurobiology

    Article Title: Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain

    doi: 10.1007/s12035-020-02276-8

    Figure Lengend Snippet: cFOS increases 4E-BP1 transcription via its interaction with Sig-1R. a Overexpression of 4E-BP1 decreases Cav2.2 in HEK cells. b Sig-1R interacts with cFOS in HEK cells. In cFOS-Myc/DDK and Sig-1R-GFP HEK co-expressing HEK cells, sample blot demonstrates the co-immunoprecipitation of those two proteins (lane 6). c Co-overexpressed Sig-1R and cFOS bind to the promoter of 4E-BP1. DNAs obtained from chromatin immunoprecipitation assays testing the binding of GFP-tagged Sig-1R (upper panel) or Myc-tagged cFOS (lower panel) to the 4E-BP1 promoter in HEK cells were examined. The precipitated DNA was amplified using specific primers for the 4E-BP1 promoter. Quantified results from the PCR products were normalized to input. Summary data are presented as means ± SEM; Student’s t test; * P < 0.05, *** P < 0.001; N = 3

    Article Snippet: Plasmid vectors used were pCMV6-CACNA1B-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC217170), pCMV6-EIF4EBP1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC201348), pCMV6-FOS-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC202597), pCMV6-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. PS100001), pCMV6-MZF1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC220791), pCMV6-OCT1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC208599), pCMV3-GFP control vector (Sino Biological, Wayne, PA; Cat. CV026), pCMV3-HA-Sec61β□ (Sino Biological, Wayne, PA; Cat. MG51753-NY), pCMV3-HA-SP3 (Sino Biological, Wayne, PA; Cat. HG13964-CY), pCMV3-Sigma-1R-GFP (Sino Biological, Wayne, PA; Cat. MG57873-ACG), pCMV3-HA control vector (Sino Biological, Wayne, PA; Cat. CV017), pcDNA3-HA-CEBPD (self-construct), pcDNA3-HA-SOX2 (self-construct), and Sigma-1R-EYFP (self-construct).

    Techniques: Over Expression, Expressing, Immunoprecipitation, Chromatin Immunoprecipitation, Binding Assay, Amplification

    Sig-1R regulates 4E-BP1 and its effect on eIF4E. a Sig-1R KO decreases the transcription and translation of 4E-BP1 in HEK cells. Sample blot demonstrates a decrease of 4E-BP1 protein in Sig-1R KO HEK cells. Summary data reveal an elevated level of protein and mRNA of 4E-BP1 in Sig-1R KO HEK cells. b Sig-1R knockout (KO) increases eIF4E and Cav2.2 mRNA binding. In the RNA-IP test, eIF4E binding to Cav2.2 5′cap region increases in Sig-1R KO HEK cells. Data represent means ± SEM; Student’s t test; Mann-Whitney test for b ; * P < 0.05, *** P < 0.001

    Journal: Molecular Neurobiology

    Article Title: Genomic Action of Sigma-1 Receptor Chaperone Relates to Neuropathic Pain

    doi: 10.1007/s12035-020-02276-8

    Figure Lengend Snippet: Sig-1R regulates 4E-BP1 and its effect on eIF4E. a Sig-1R KO decreases the transcription and translation of 4E-BP1 in HEK cells. Sample blot demonstrates a decrease of 4E-BP1 protein in Sig-1R KO HEK cells. Summary data reveal an elevated level of protein and mRNA of 4E-BP1 in Sig-1R KO HEK cells. b Sig-1R knockout (KO) increases eIF4E and Cav2.2 mRNA binding. In the RNA-IP test, eIF4E binding to Cav2.2 5′cap region increases in Sig-1R KO HEK cells. Data represent means ± SEM; Student’s t test; Mann-Whitney test for b ; * P < 0.05, *** P < 0.001

    Article Snippet: Plasmid vectors used were pCMV6-CACNA1B-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC217170), pCMV6-EIF4EBP1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC201348), pCMV6-FOS-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC202597), pCMV6-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. PS100001), pCMV6-MZF1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC220791), pCMV6-OCT1-myc/DDK (Origene Technologies Inc., Rockville, MD; Cat. RC208599), pCMV3-GFP control vector (Sino Biological, Wayne, PA; Cat. CV026), pCMV3-HA-Sec61β□ (Sino Biological, Wayne, PA; Cat. MG51753-NY), pCMV3-HA-SP3 (Sino Biological, Wayne, PA; Cat. HG13964-CY), pCMV3-Sigma-1R-GFP (Sino Biological, Wayne, PA; Cat. MG57873-ACG), pCMV3-HA control vector (Sino Biological, Wayne, PA; Cat. CV017), pcDNA3-HA-CEBPD (self-construct), pcDNA3-HA-SOX2 (self-construct), and Sigma-1R-EYFP (self-construct).

    Techniques: Knock-Out, Binding Assay, MANN-WHITNEY