genex plus transfection reagent  (ATCC)


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    ATCC genex plus transfection reagent
    Genex Plus Transfection Reagent, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    genex plus transfection reagent  (ATCC)


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    ATCC genex plus transfection reagent
    Genex Plus Transfection Reagent, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    genexplus transfection reagent  (ATCC)


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    ATCC genexplus transfection reagent
    Genexplus Transfection Reagent, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    genexplus  (ATCC)


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    ATCC genexplus
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    porcine kidney 15 pk15  (ATCC)


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    ATCC porcine kidney 15 pk15
    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine <t>kidney-15</t> <t>(PK15)</t> and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    Porcine Kidney 15 Pk15, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs"

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    Journal: Biology

    doi: 10.3390/biology11010135

    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    Figure Legend Snippet: Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .

    Techniques Used: Expressing, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Immunoprecipitation, Polymerase Chain Reaction, Amplification, Western Blot

    Transcription factor IRF1 promoted the transcription activity of DRD2 gene. ( A ) The IRF1 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized to GAPDH. ( B ) DRD2 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. ( C ) Analysis of the binding region of IRF1; pcIRF1 and pGL3-promoter-WT or pGL3-promoter-MUT vectors were co-transfected into PK15 cells. The pcDNA3.1 vector was applied as a vector control. ( D ) Immunofluorescence staining of MAP2 or TUJ1 expression in the PNCs. Nuclei were stained using DAPI. ( E , F ) RT-qPCR analyzed the IRF1 knockdown efficiency and DRD2 gene mRNA expression level. PK15 and PNCs were transfected with two different siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized using GAPDH. ( G , H ) The IRF1 knockdown efficiency and the protein level of the DRD2 gene were determined by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .
    Figure Legend Snippet: Transcription factor IRF1 promoted the transcription activity of DRD2 gene. ( A ) The IRF1 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized to GAPDH. ( B ) DRD2 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. ( C ) Analysis of the binding region of IRF1; pcIRF1 and pGL3-promoter-WT or pGL3-promoter-MUT vectors were co-transfected into PK15 cells. The pcDNA3.1 vector was applied as a vector control. ( D ) Immunofluorescence staining of MAP2 or TUJ1 expression in the PNCs. Nuclei were stained using DAPI. ( E , F ) RT-qPCR analyzed the IRF1 knockdown efficiency and DRD2 gene mRNA expression level. PK15 and PNCs were transfected with two different siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized using GAPDH. ( G , H ) The IRF1 knockdown efficiency and the protein level of the DRD2 gene were determined by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Techniques Used: Activity Assay, Expressing, Transfection, Binding Assay, Plasmid Preparation, Immunofluorescence, Staining, Quantitative RT-PCR, Negative Control, Western Blot, Infection

    Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .
    Figure Legend Snippet: Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Techniques Used: Expressing, Mutagenesis, Binding Assay, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Construct, Immunoprecipitation, Amplification, Western Blot

    IRF2 as a transcription repressor downregulated the DRD2 gene expression. ( A , B ) IRF2 mRNA expression in PK15 cells which transfected with pcIRF2 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. IRF2 overexpression diminished DRD2 transcription in PK15 cells. ( C ) Analysis of the binding region of IRF2 when co-transfected pcIRF2 and pGL3-DRD2-WT or pGL3-DRD2-MUT vectors into PK15 cells; the pcDNA3.1 vector was applied as a vector control. ( D , E ) The IRF2 knockdown efficiency and the expression of DRD2 gene were detected by RT-qPCR analysis. PK15 and PNCs were infected with two different IRF2 siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized to GAPDH. ( F , G ) IRF2 knockdown efficiency and the protein level of DRD2 gene were detected by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .
    Figure Legend Snippet: IRF2 as a transcription repressor downregulated the DRD2 gene expression. ( A , B ) IRF2 mRNA expression in PK15 cells which transfected with pcIRF2 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. IRF2 overexpression diminished DRD2 transcription in PK15 cells. ( C ) Analysis of the binding region of IRF2 when co-transfected pcIRF2 and pGL3-DRD2-WT or pGL3-DRD2-MUT vectors into PK15 cells; the pcDNA3.1 vector was applied as a vector control. ( D , E ) The IRF2 knockdown efficiency and the expression of DRD2 gene were detected by RT-qPCR analysis. PK15 and PNCs were infected with two different IRF2 siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized to GAPDH. ( F , G ) IRF2 knockdown efficiency and the protein level of DRD2 gene were detected by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Techniques Used: Expressing, Transfection, Over Expression, Binding Assay, Plasmid Preparation, Quantitative RT-PCR, Infection, Negative Control, Western Blot

    293t  (ATCC)


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    ATCC 293t
    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into <t>293T</t> cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    293t, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs"

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    Journal: Biology

    doi: 10.3390/biology11010135

    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    Figure Legend Snippet: Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .

    Techniques Used: Expressing, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Immunoprecipitation, Polymerase Chain Reaction, Amplification, Western Blot

    Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .
    Figure Legend Snippet: Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Techniques Used: Expressing, Mutagenesis, Binding Assay, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Construct, Immunoprecipitation, Amplification, Western Blot

    genexplus  (ATCC)


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    ATCC genexplus
    Genexplus, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    genex plus transfection reagent atcc atcc acs 4004 series scm5chip  (ATCC)


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    ATCC genex plus transfection reagent atcc atcc acs 4004 series scm5chip
    Genex Plus Transfection Reagent Atcc Atcc Acs 4004 Series Scm5chip, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    genex plus transfection reagent  (ATCC)


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    ATCC genex plus transfection reagent
    Genex Plus Transfection Reagent, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    genexplus  (ATCC)


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    ATCC genexplus
    Genexplus, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    atcc acs  (ATCC)


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    ATCC atcc acs
    Atcc Acs, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC genex plus transfection reagent
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    ATCC porcine kidney 15 pk15
    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine <t>kidney-15</t> <t>(PK15)</t> and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    Porcine Kidney 15 Pk15, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    293t  (ATCC)
    99
    ATCC 293t
    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into <t>293T</t> cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    293t, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC genex plus transfection reagent atcc atcc acs 4004 series scm5chip
    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into <t>293T</t> cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    Genex Plus Transfection Reagent Atcc Atcc Acs 4004 Series Scm5chip, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC atcc acs
    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into <t>293T</t> cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .
    Atcc Acs, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .

    Journal: Biology

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    doi: 10.3390/biology11010135

    Figure Lengend Snippet: Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .

    Article Snippet: Porcine kidney-15 (PK15) (ATCC ® ACS-4004 ™ ) and 293T (ATCC ® ACS-4004 ™ ) cells were cultured in high-glucose medium with 10% fetal bovine serum (FBS) (10%FBS + 90%DMEM) (FBS, Gibco) (DMEM/High, Gibco) at 37 °C and 5% CO 2 .

    Techniques: Expressing, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Immunoprecipitation, Polymerase Chain Reaction, Amplification, Western Blot

    Transcription factor IRF1 promoted the transcription activity of DRD2 gene. ( A ) The IRF1 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized to GAPDH. ( B ) DRD2 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. ( C ) Analysis of the binding region of IRF1; pcIRF1 and pGL3-promoter-WT or pGL3-promoter-MUT vectors were co-transfected into PK15 cells. The pcDNA3.1 vector was applied as a vector control. ( D ) Immunofluorescence staining of MAP2 or TUJ1 expression in the PNCs. Nuclei were stained using DAPI. ( E , F ) RT-qPCR analyzed the IRF1 knockdown efficiency and DRD2 gene mRNA expression level. PK15 and PNCs were transfected with two different siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized using GAPDH. ( G , H ) The IRF1 knockdown efficiency and the protein level of the DRD2 gene were determined by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Journal: Biology

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    doi: 10.3390/biology11010135

    Figure Lengend Snippet: Transcription factor IRF1 promoted the transcription activity of DRD2 gene. ( A ) The IRF1 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized to GAPDH. ( B ) DRD2 mRNA expression in PK15 cells which transfected with pcIRF1 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. ( C ) Analysis of the binding region of IRF1; pcIRF1 and pGL3-promoter-WT or pGL3-promoter-MUT vectors were co-transfected into PK15 cells. The pcDNA3.1 vector was applied as a vector control. ( D ) Immunofluorescence staining of MAP2 or TUJ1 expression in the PNCs. Nuclei were stained using DAPI. ( E , F ) RT-qPCR analyzed the IRF1 knockdown efficiency and DRD2 gene mRNA expression level. PK15 and PNCs were transfected with two different siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized using GAPDH. ( G , H ) The IRF1 knockdown efficiency and the protein level of the DRD2 gene were determined by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Article Snippet: Porcine kidney-15 (PK15) (ATCC ® ACS-4004 ™ ) and 293T (ATCC ® ACS-4004 ™ ) cells were cultured in high-glucose medium with 10% fetal bovine serum (FBS) (10%FBS + 90%DMEM) (FBS, Gibco) (DMEM/High, Gibco) at 37 °C and 5% CO 2 .

    Techniques: Activity Assay, Expressing, Transfection, Binding Assay, Plasmid Preparation, Immunofluorescence, Staining, Quantitative RT-PCR, Negative Control, Western Blot, Infection

    Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Journal: Biology

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    doi: 10.3390/biology11010135

    Figure Lengend Snippet: Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Article Snippet: Porcine kidney-15 (PK15) (ATCC ® ACS-4004 ™ ) and 293T (ATCC ® ACS-4004 ™ ) cells were cultured in high-glucose medium with 10% fetal bovine serum (FBS) (10%FBS + 90%DMEM) (FBS, Gibco) (DMEM/High, Gibco) at 37 °C and 5% CO 2 .

    Techniques: Expressing, Mutagenesis, Binding Assay, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Construct, Immunoprecipitation, Amplification, Western Blot

    IRF2 as a transcription repressor downregulated the DRD2 gene expression. ( A , B ) IRF2 mRNA expression in PK15 cells which transfected with pcIRF2 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. IRF2 overexpression diminished DRD2 transcription in PK15 cells. ( C ) Analysis of the binding region of IRF2 when co-transfected pcIRF2 and pGL3-DRD2-WT or pGL3-DRD2-MUT vectors into PK15 cells; the pcDNA3.1 vector was applied as a vector control. ( D , E ) The IRF2 knockdown efficiency and the expression of DRD2 gene were detected by RT-qPCR analysis. PK15 and PNCs were infected with two different IRF2 siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized to GAPDH. ( F , G ) IRF2 knockdown efficiency and the protein level of DRD2 gene were detected by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Journal: Biology

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    doi: 10.3390/biology11010135

    Figure Lengend Snippet: IRF2 as a transcription repressor downregulated the DRD2 gene expression. ( A , B ) IRF2 mRNA expression in PK15 cells which transfected with pcIRF2 or pcDNA3.1(+). The mRNA levels were normalized using GAPDH. IRF2 overexpression diminished DRD2 transcription in PK15 cells. ( C ) Analysis of the binding region of IRF2 when co-transfected pcIRF2 and pGL3-DRD2-WT or pGL3-DRD2-MUT vectors into PK15 cells; the pcDNA3.1 vector was applied as a vector control. ( D , E ) The IRF2 knockdown efficiency and the expression of DRD2 gene were detected by RT-qPCR analysis. PK15 and PNCs were infected with two different IRF2 siRNAs and negative control (NC) (2.5 μL) by lip3000 for 24 h. The mRNA levels were normalized to GAPDH. ( F , G ) IRF2 knockdown efficiency and the protein level of DRD2 gene were detected by western blotting analysis. PK15 and PNCs were infected with the siRNA and negative control (NC) (5 μL) by lip3000 for 48 h. Data were presented as mean ± SE of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Article Snippet: Porcine kidney-15 (PK15) (ATCC ® ACS-4004 ™ ) and 293T (ATCC ® ACS-4004 ™ ) cells were cultured in high-glucose medium with 10% fetal bovine serum (FBS) (10%FBS + 90%DMEM) (FBS, Gibco) (DMEM/High, Gibco) at 37 °C and 5% CO 2 .

    Techniques: Expressing, Transfection, Over Expression, Binding Assay, Plasmid Preparation, Quantitative RT-PCR, Infection, Negative Control, Western Blot

    Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .

    Journal: Biology

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    doi: 10.3390/biology11010135

    Figure Lengend Snippet: Expression profile of porcine DRD2 and IRF1 gene, and site-directed mutagenesis of IRF1 binding site in the DRD2 promoter. ( A , B ) Expression characteristics of porcine DRD2 and IRF1 gene at different tissues. ( C ) Site-directed mutation schematic diagram of the predicted IRF1 binding site in the DRD2 promoter. ( D ) Site-directed mutation of IRF1 binding site of DRD2 gene by luciferase activity assay. Wild-type (WT) or mutant (MUT) of the DRD2 promoter were transfected into 293T cells, respectively. The pGMLR-TK luciferase reporter vector was applied as an internal control, and the pGL3-Basic vector was used as a negative control, the pGL3-control vector was used as a positive control. ( E ) Binding of IRF1 on the DRD2 promoter was demonstrated using ChIP assays in porcine kidney-15 (PK15) and neuronal cells (PNCs). After immunoprecipitation, identified the IRF1 binding site by polymerase chain reaction (PCR) amplification. Input was total fragmented DNA. Precipitated chromatin with normal Immunoglobulin G (IgG) was applied as the negative control. Data were presented as means ± standard errors (SE) of three replicates. ** p < 0.01. The uncropped western blot figures can be accessed in .

    Article Snippet: Porcine kidney-15 (PK15) (ATCC ® ACS-4004 ™ ) and 293T (ATCC ® ACS-4004 ™ ) cells were cultured in high-glucose medium with 10% fetal bovine serum (FBS) (10%FBS + 90%DMEM) (FBS, Gibco) (DMEM/High, Gibco) at 37 °C and 5% CO 2 .

    Techniques: Expressing, Mutagenesis, Binding Assay, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Immunoprecipitation, Polymerase Chain Reaction, Amplification, Western Blot

    Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Journal: Biology

    Article Title: Functionally Antagonistic Transcription Factors IRF1 and IRF2 Regulate the Transcription of the Dopamine Receptor D2 Gene Associated with Aggressive Behavior of Weaned Pigs

    doi: 10.3390/biology11010135

    Figure Lengend Snippet: Expression profile of porcine IRF2 gene and site-directed mutagenesis of IRF2 binding site in the DRD2 transcriptional suppression region. ( A ) Expression characteristics of porcine IRF2 gene at different tissues by RT-qPCR analysis. ( B ) Site-directed mutation schematic diagram of the predicted IRF2 binding site in the DRD2 promoter. ( C ) Site-directed mutation of IRF2 binding site of DRD2 gene by luciferase activity assay. Wild-type or mutant of the DRD2 promoter luciferase reporter vectors were transfected into 293T cells, respectively. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( D ) Luciferase reporter gene assays of porcine DRD2 alleles containing rs1110730503 (−915A/T). Two DRD2 genotype luciferase reporter vectors were constructed and transfected into 293T cells. The pGL3-Basic vector was used as a negative control, and the pGL3-control vector was used as a positive control. ( E ) The IRF2 binding sits on the DRD2 gene were identified using ChIP assays in PK15 and PNCs. After immunoprecipitation, the IRF2 binding sites were demonstrated by PCR amplification. Input was total fragmented DNA. Precipitated chromatin with normal IgG was applied as the negative control. Data were presented as mean ± SE. of three replicates. * p < 0.05, ** p < 0.01. The uncropped western blot figures can be accessed in .

    Article Snippet: Porcine kidney-15 (PK15) (ATCC ® ACS-4004 ™ ) and 293T (ATCC ® ACS-4004 ™ ) cells were cultured in high-glucose medium with 10% fetal bovine serum (FBS) (10%FBS + 90%DMEM) (FBS, Gibco) (DMEM/High, Gibco) at 37 °C and 5% CO 2 .

    Techniques: Expressing, Mutagenesis, Binding Assay, Quantitative RT-PCR, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Positive Control, Construct, Immunoprecipitation, Amplification, Western Blot