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    Name:
    GR Antibody
    Description:
    Anti GR Antibody G 5 is a mouse monoclonal IgG2b kappa light chain GR antibody provided at 200 µg ml raised against amino acids 121 420 of GR of human origin Anti GR Antibody G 5 is recommended for detection of GR α and GR β of mouse rat and human origin by WB IP IF IHC P and ELISA Anti GR Antibody G 5 is available conjugated to agarose for IP HRP for WB IHC P and ELISA and to either phycoerythrin or FITC for IF IHC P and FCM also available conjugated to Alexa Fluor 488 Alexa Fluor 546 Alexa Fluor 594 or Alexa Fluor 647 for WB RGB IF IHC P and FCM and for use with RGB fluorescent imaging systems such as iBright FL1000 FluorChem Typhoon Azure and other comparable systems also available conjugated to Alexa Fluor 680 or Alexa Fluor 790 for WB NIR IF and FCM for use with Near Infrared NIR detection systems such as LI COR Odyssey iBright FL1000 FluorChem Typhoon Azure and other comparable systems TransCruz reagent for Gel Supershift and ChIP applications sc 393232 X 200 µg 0 1 ml Contact our Technical Service Department or your local Distributor for more information on how to receive a FREE 10 µg sample of GR G 5 sc 393232
    Catalog Number:
    SC-393232
    Price:
    None
    Category:
    Antibodies Primary Antibodies and ImmunoCruz Conjugates Steroid Receptors GR Antibodies GR Antibody G 5
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    Structured Review

    Santa Cruz Biotechnology figr
    Interaction of <t>FKBP51</t> with GRα and PPARγ. GRα and PPARγ transfected COS-7 cells were adsorbed to protein G-Sepharose using the <t>FiGR</t> monoclonal antibody to GRα (I), PPARγ monoclonal antibody (I), or nonimmune
    Anti GR Antibody G 5 is a mouse monoclonal IgG2b kappa light chain GR antibody provided at 200 µg ml raised against amino acids 121 420 of GR of human origin Anti GR Antibody G 5 is recommended for detection of GR α and GR β of mouse rat and human origin by WB IP IF IHC P and ELISA Anti GR Antibody G 5 is available conjugated to agarose for IP HRP for WB IHC P and ELISA and to either phycoerythrin or FITC for IF IHC P and FCM also available conjugated to Alexa Fluor 488 Alexa Fluor 546 Alexa Fluor 594 or Alexa Fluor 647 for WB RGB IF IHC P and FCM and for use with RGB fluorescent imaging systems such as iBright FL1000 FluorChem Typhoon Azure and other comparable systems also available conjugated to Alexa Fluor 680 or Alexa Fluor 790 for WB NIR IF and FCM for use with Near Infrared NIR detection systems such as LI COR Odyssey iBright FL1000 FluorChem Typhoon Azure and other comparable systems TransCruz reagent for Gel Supershift and ChIP applications sc 393232 X 200 µg 0 1 ml Contact our Technical Service Department or your local Distributor for more information on how to receive a FREE 10 µg sample of GR G 5 sc 393232
    https://www.bioz.com/result/figr/product/Santa Cruz Biotechnology
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    figr - by Bioz Stars, 2021-09
    95/100 stars

    Images

    1) Product Images from "FKBP51 Reciprocally Regulates GRα and PPARγ Activation via the Akt-p38 Pathway"

    Article Title: FKBP51 Reciprocally Regulates GRα and PPARγ Activation via the Akt-p38 Pathway

    Journal: Molecular Endocrinology

    doi: 10.1210/me.2014-1023

    Interaction of FKBP51 with GRα and PPARγ. GRα and PPARγ transfected COS-7 cells were adsorbed to protein G-Sepharose using the FiGR monoclonal antibody to GRα (I), PPARγ monoclonal antibody (I), or nonimmune
    Figure Legend Snippet: Interaction of FKBP51 with GRα and PPARγ. GRα and PPARγ transfected COS-7 cells were adsorbed to protein G-Sepharose using the FiGR monoclonal antibody to GRα (I), PPARγ monoclonal antibody (I), or nonimmune

    Techniques Used: Transfection

    2) Product Images from "Analysis of FK506, timcodar (VX-853) and FKBP51 and FKBP52 chaperones in control of glucocorticoid receptor activity and phosphorylation"

    Article Title: Analysis of FK506, timcodar (VX-853) and FKBP51 and FKBP52 chaperones in control of glucocorticoid receptor activity and phosphorylation

    Journal: Pharmacology Research & Perspectives

    doi: 10.1002/prp2.76

    FKBP51 and FKBP52 reciprocally control GR activity and phosphorylation. (A) Western blot analysis of whole cell extracts from WT, FKBP51-KO, and FKBP52-KO MEF cells demonstrating a complete lack of FKBP51 and FKBP52 in the KO cells. GAPDH was used as loading control. (B) qRT-PCR analysis of SGK and GILZ expression in WT, FKBP51-KO, and FKBP52-KO MEF cells following treatment with 100 μ mol/L Dex for 2 h. Transcript expression was normalized to 18S mRNA. Data represent the mean ± SEM of three independent experiments, assayed in duplicate. *versus WT control, † KO versus WT. (C) Whole cell extracts of WT, FKBP51-KO and FKBP52-KO MEF cells treated with or without Dex for 1 h were analyzed by Western blotting with antibodies specific to phospho-serines 212, 220, and 234 of mouse GR. FiGR antibody was used to detect total GR. Extracts from COS-7 cells lacking GR were used as negative controls (neg ctrl). Quantitation of GR bands was performed by infrared spectrophotometry. Phospho-GR (pGR) signals were normalized to total GR at each condition. Data represent the mean ± SEM of three independent experiments. Significant differences in transcript expression or protein levels are indicated as follows: * P
    Figure Legend Snippet: FKBP51 and FKBP52 reciprocally control GR activity and phosphorylation. (A) Western blot analysis of whole cell extracts from WT, FKBP51-KO, and FKBP52-KO MEF cells demonstrating a complete lack of FKBP51 and FKBP52 in the KO cells. GAPDH was used as loading control. (B) qRT-PCR analysis of SGK and GILZ expression in WT, FKBP51-KO, and FKBP52-KO MEF cells following treatment with 100 μ mol/L Dex for 2 h. Transcript expression was normalized to 18S mRNA. Data represent the mean ± SEM of three independent experiments, assayed in duplicate. *versus WT control, † KO versus WT. (C) Whole cell extracts of WT, FKBP51-KO and FKBP52-KO MEF cells treated with or without Dex for 1 h were analyzed by Western blotting with antibodies specific to phospho-serines 212, 220, and 234 of mouse GR. FiGR antibody was used to detect total GR. Extracts from COS-7 cells lacking GR were used as negative controls (neg ctrl). Quantitation of GR bands was performed by infrared spectrophotometry. Phospho-GR (pGR) signals were normalized to total GR at each condition. Data represent the mean ± SEM of three independent experiments. Significant differences in transcript expression or protein levels are indicated as follows: * P

    Techniques Used: Activity Assay, Western Blot, Quantitative RT-PCR, Expressing, Quantitation Assay, Spectrophotometry

    Related Articles

    Chromatin Immunoprecipitation:

    Article Title: The long noncoding RNA HOTAIRM1 controlled by AML1 enhances glucocorticoid resistance by activating RHOA/ROCK1 pathway through suppressing ARHGAP18
    Article Snippet: .. And ChIP was performed by co-precipitating the DNA/protein complexes with a GR antibody (sc-393232) (Santa Cruz Biotechnology, Texas, USA) and a rabbit control IgG (Santa Cruz Biotechnology, Texas, USA). ..

    Incubation:

    Article Title: Glucocorticoid receptor signaling in ventral tegmental area neurons increases the rewarding value of a high-fat diet in mice
    Article Snippet: .. Next, the sections were incubated with anti-DAT antibody (1:500; Merck Millipore) or anti-GR (1:500; Santa Cruz Biotechnology, CA, USA) overnight at 4 °C. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Ahi1 regulates the nuclear translocation of glucocorticoid receptor to modulate stress response
    Article Snippet: .. Antibodies and reagents Ahi1 Rabbit Antibody and Hap1 Guinea pigs Antibody were described previously ; GR antibody (sc-56851) and His-probe antibody (sc-53073) were obtained from Santa-Cruz Biotechnology Inc. (Santa-Cruz, CA, USA); LSD-1 (#2139), and Histone-3 (#4499) antibodies were from Cell Signaling Technology, Inc., (Danver, MA, USA);ubiquitin antibody was from Abcam, (ab134953,Cambridge, MA, USA), Beta-tubulin antibody, anti-beta-actin antibody, imipramine (IM), mifepristone (RU 38486), fluoxetine, MG132, and dexamethasone acetate (Dex) were from Sigma-Aldrich Company (St. Louis, Missouri, USA); RPMI-1640 and FBS (Gibco Company, MD, USA); ECL chemiluminescence system was from Thermo Company (West Chester, PA, USA); Mouse corticosterone ELISA kit was purchased from Cusabio Biological Engineering Co. LTD (Baltimore, MD, USA); Transcriptor First Strand cDNA Synthesis Kit and 2XSYBR Green PCR Master Mix (Roche, Germany); Lipofectamine 2000 was bought from Invitrogen Corporation (San Diego, CA, USA). ..

    Polymerase Chain Reaction:

    Article Title: Ahi1 regulates the nuclear translocation of glucocorticoid receptor to modulate stress response
    Article Snippet: .. Antibodies and reagents Ahi1 Rabbit Antibody and Hap1 Guinea pigs Antibody were described previously ; GR antibody (sc-56851) and His-probe antibody (sc-53073) were obtained from Santa-Cruz Biotechnology Inc. (Santa-Cruz, CA, USA); LSD-1 (#2139), and Histone-3 (#4499) antibodies were from Cell Signaling Technology, Inc., (Danver, MA, USA);ubiquitin antibody was from Abcam, (ab134953,Cambridge, MA, USA), Beta-tubulin antibody, anti-beta-actin antibody, imipramine (IM), mifepristone (RU 38486), fluoxetine, MG132, and dexamethasone acetate (Dex) were from Sigma-Aldrich Company (St. Louis, Missouri, USA); RPMI-1640 and FBS (Gibco Company, MD, USA); ECL chemiluminescence system was from Thermo Company (West Chester, PA, USA); Mouse corticosterone ELISA kit was purchased from Cusabio Biological Engineering Co. LTD (Baltimore, MD, USA); Transcriptor First Strand cDNA Synthesis Kit and 2XSYBR Green PCR Master Mix (Roche, Germany); Lipofectamine 2000 was bought from Invitrogen Corporation (San Diego, CA, USA). ..

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  • 95
    Santa Cruz Biotechnology rabbit polyclonal antiserum against gr
    Activation of the ERK or the JNK pathway inhibits GR transcriptional activation. ( A ) Selective activation of ERK and JNK in HeLa cells by f-sos and racQ61L. pRK5-HA ERK (1.5 μg), pRK5-M2 JNK (1.5 μg), along with pRK5-HA f-sos (4 μg) or pCMV5-racQ61L (4 μg) or an empty pRK5 vector (4 μg), where indicated, was transfected into HeLa cells (2.5 × 10 5 cells/60-mm dish) using Lipofectamine. Transfected cells were treated with 100 nM Dex where indicated and harvested 8 h later. To visualize HA ERK and HA f-sos, WCE from ERK-transfected cells were immunoblotted with anti-HA mouse monoclonal antibody (left panel). FLAG-tagged JNK was immunoprecipitated from WCE by using the FLAG-specific mouse monoclonal antibody M2 and used in vitro to phosphorylate recombinant purified GST–c-Jun. Phosphorylated proteins were separated on 10% SDS/polyacrylamide gels, stained with Coomassie blue to verify equal amount of GST–c-Jun in each lane, and autoradiographed (right panel). ( B ) Inhibition of GR transcriptional enhancement by activated ERK and JNK. HeLa cells were transfected as described in A ) and normalized to β-Gal activity. ( C ) Level of GR protein is unaffected by JNK or ERK activation. WCE were prepared from a parallel set of transfected cells. Equal amounts of total protein (50 μg/lane) were separated by SDS/PAGE, transferred to Immobilon paper, probed with a GR-specific <t>polyclonal</t> antibody, and visualized by enhanced chemiluminescence.
    Rabbit Polyclonal Antiserum Against Gr, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal antiserum against gr/product/Santa Cruz Biotechnology
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal antiserum against gr - by Bioz Stars, 2021-09
    95/100 stars
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    95
    Santa Cruz Biotechnology antibodies against grα
    Skeletal muscle conditioned medium inhibited the adipogenesis by reducing <t>GRα</t> expression. The subcutaneous pre-adipocytes were cultured with skeletal muscle conditioned medium (MCM) or ordinary medium for 3 days, then the cells were collected for RNA, protein isolation and flow cytometry assay. ( a ) The mRNA expression of GRα and E1-C, E1-H was detected by RT-qPCR, n = 6 per group. ( b ) Expression of GRα protein was detected by Western blot (left) and displayed as column charts after quantification (right), n = 3 per group, * P
    Antibodies Against Grα, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies against grα/product/Santa Cruz Biotechnology
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibodies against grα - by Bioz Stars, 2021-09
    95/100 stars
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    86
    Santa Cruz Biotechnology polyclonal anti gr
    Dex increases the abundance of nuclear GR. Western blot analysis using a <t>polyclonal</t> anti-GR antibody were performed on nuclear extracts of A549 cells. ( A ) Cells were exposed to vehicle (−) or 100 nM Dex (+) for 24 h; 7 or 14 μg of nuclear protein (NP) were analyzed. ( B ) Cells were exposed to 100 nM Dex for the times indicated. Ten micrograms of NP were evaluated for GR and actin protein for each time point. Results are shown as the relative fold induction of Dex-induced nuclear GR levels normalized to actin levels, expressed as the mean ± SE. Experiments were performed in triplicate on three separate occasions. * P
    Polyclonal Anti Gr, supplied by Santa Cruz Biotechnology, 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/polyclonal anti gr/product/Santa Cruz Biotechnology
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    polyclonal anti gr - by Bioz Stars, 2021-09
    86/100 stars
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    Image Search Results


    Activation of the ERK or the JNK pathway inhibits GR transcriptional activation. ( A ) Selective activation of ERK and JNK in HeLa cells by f-sos and racQ61L. pRK5-HA ERK (1.5 μg), pRK5-M2 JNK (1.5 μg), along with pRK5-HA f-sos (4 μg) or pCMV5-racQ61L (4 μg) or an empty pRK5 vector (4 μg), where indicated, was transfected into HeLa cells (2.5 × 10 5 cells/60-mm dish) using Lipofectamine. Transfected cells were treated with 100 nM Dex where indicated and harvested 8 h later. To visualize HA ERK and HA f-sos, WCE from ERK-transfected cells were immunoblotted with anti-HA mouse monoclonal antibody (left panel). FLAG-tagged JNK was immunoprecipitated from WCE by using the FLAG-specific mouse monoclonal antibody M2 and used in vitro to phosphorylate recombinant purified GST–c-Jun. Phosphorylated proteins were separated on 10% SDS/polyacrylamide gels, stained with Coomassie blue to verify equal amount of GST–c-Jun in each lane, and autoradiographed (right panel). ( B ) Inhibition of GR transcriptional enhancement by activated ERK and JNK. HeLa cells were transfected as described in A ) and normalized to β-Gal activity. ( C ) Level of GR protein is unaffected by JNK or ERK activation. WCE were prepared from a parallel set of transfected cells. Equal amounts of total protein (50 μg/lane) were separated by SDS/PAGE, transferred to Immobilon paper, probed with a GR-specific polyclonal antibody, and visualized by enhanced chemiluminescence.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Antagonism of glucocorticoid receptor transcriptional activation by the c-Jun N-terminal kinase

    doi:

    Figure Lengend Snippet: Activation of the ERK or the JNK pathway inhibits GR transcriptional activation. ( A ) Selective activation of ERK and JNK in HeLa cells by f-sos and racQ61L. pRK5-HA ERK (1.5 μg), pRK5-M2 JNK (1.5 μg), along with pRK5-HA f-sos (4 μg) or pCMV5-racQ61L (4 μg) or an empty pRK5 vector (4 μg), where indicated, was transfected into HeLa cells (2.5 × 10 5 cells/60-mm dish) using Lipofectamine. Transfected cells were treated with 100 nM Dex where indicated and harvested 8 h later. To visualize HA ERK and HA f-sos, WCE from ERK-transfected cells were immunoblotted with anti-HA mouse monoclonal antibody (left panel). FLAG-tagged JNK was immunoprecipitated from WCE by using the FLAG-specific mouse monoclonal antibody M2 and used in vitro to phosphorylate recombinant purified GST–c-Jun. Phosphorylated proteins were separated on 10% SDS/polyacrylamide gels, stained with Coomassie blue to verify equal amount of GST–c-Jun in each lane, and autoradiographed (right panel). ( B ) Inhibition of GR transcriptional enhancement by activated ERK and JNK. HeLa cells were transfected as described in A ) and normalized to β-Gal activity. ( C ) Level of GR protein is unaffected by JNK or ERK activation. WCE were prepared from a parallel set of transfected cells. Equal amounts of total protein (50 μg/lane) were separated by SDS/PAGE, transferred to Immobilon paper, probed with a GR-specific polyclonal antibody, and visualized by enhanced chemiluminescence.

    Article Snippet: For Western blotting, protein extracts were fractionated on 10% SDS/polyacrylamide gels, transferred to Immobilon paper (Millipore), and probed with mouse monoclonal antibodies against hemagglutinin (HA) (Boehringer Mannheim), or FLAG (M2, Kodak), or with a rabbit polyclonal antiserum against GR (Santa Cruz Biotechnology).

    Techniques: Activation Assay, Plasmid Preparation, Transfection, Immunoprecipitation, In Vitro, Recombinant, Purification, Staining, Inhibition, Activity Assay, SDS Page

    Activation of the JNK but not ERK pathway in vivo increases GR phosphorylation. HeLa cells (1 × 10 6 ) were transiently transfected in 100-mm dishes with pCMV-GR S224A S232A (2.3 μg) and ( A ) pRK5-M2 JNK (4.5 μg) with pCMV5-racQ61L (12 μg) or an empty pCMV5 vector (12 μg) or ( B ) pRK5-HA ERK (4.5 μg) with pRK5-HA f-sos (12 μg) or an empty pRK5 vector (12 μg) by using Lipofectamine. After 18 h, GR was metabolically labeled for 2 h with 1 mCi/ml of [ 32 P]orthophosphate in the presence of 100 nM Dex. In vivo labeled receptor was immunoprecipitated from cell lysates with the rat GR-specific monoclonal antibody, BuGR2. Immunoprecipitates were separated on 9% SDS/PAGE gels, transferred to Immobilon membrane, and exposed to film to visualize the phosphorylated GR ( Upper ). To normalize to GR protein expression, Western blotting of the same membrane with anti-GR rabbit polyclonal antibody was performed ( Lower ). The densitometric value obtained in the absence of cotransfected racQ61L ( A ) or f-sos ( B ) is arbitrarily set as 1.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Antagonism of glucocorticoid receptor transcriptional activation by the c-Jun N-terminal kinase

    doi:

    Figure Lengend Snippet: Activation of the JNK but not ERK pathway in vivo increases GR phosphorylation. HeLa cells (1 × 10 6 ) were transiently transfected in 100-mm dishes with pCMV-GR S224A S232A (2.3 μg) and ( A ) pRK5-M2 JNK (4.5 μg) with pCMV5-racQ61L (12 μg) or an empty pCMV5 vector (12 μg) or ( B ) pRK5-HA ERK (4.5 μg) with pRK5-HA f-sos (12 μg) or an empty pRK5 vector (12 μg) by using Lipofectamine. After 18 h, GR was metabolically labeled for 2 h with 1 mCi/ml of [ 32 P]orthophosphate in the presence of 100 nM Dex. In vivo labeled receptor was immunoprecipitated from cell lysates with the rat GR-specific monoclonal antibody, BuGR2. Immunoprecipitates were separated on 9% SDS/PAGE gels, transferred to Immobilon membrane, and exposed to film to visualize the phosphorylated GR ( Upper ). To normalize to GR protein expression, Western blotting of the same membrane with anti-GR rabbit polyclonal antibody was performed ( Lower ). The densitometric value obtained in the absence of cotransfected racQ61L ( A ) or f-sos ( B ) is arbitrarily set as 1.

    Article Snippet: For Western blotting, protein extracts were fractionated on 10% SDS/polyacrylamide gels, transferred to Immobilon paper (Millipore), and probed with mouse monoclonal antibodies against hemagglutinin (HA) (Boehringer Mannheim), or FLAG (M2, Kodak), or with a rabbit polyclonal antiserum against GR (Santa Cruz Biotechnology).

    Techniques: Activation Assay, In Vivo, Transfection, Plasmid Preparation, Metabolic Labelling, Labeling, Immunoprecipitation, SDS Page, Expressing, Western Blot

    Skeletal muscle conditioned medium inhibited the adipogenesis by reducing GRα expression. The subcutaneous pre-adipocytes were cultured with skeletal muscle conditioned medium (MCM) or ordinary medium for 3 days, then the cells were collected for RNA, protein isolation and flow cytometry assay. ( a ) The mRNA expression of GRα and E1-C, E1-H was detected by RT-qPCR, n = 6 per group. ( b ) Expression of GRα protein was detected by Western blot (left) and displayed as column charts after quantification (right), n = 3 per group, * P

    Journal: Scientific Reports

    Article Title: Muscle-specific downregulation of GR levels inhibits adipogenesis in porcine intramuscular adipocyte tissue

    doi: 10.1038/s41598-017-00615-9

    Figure Lengend Snippet: Skeletal muscle conditioned medium inhibited the adipogenesis by reducing GRα expression. The subcutaneous pre-adipocytes were cultured with skeletal muscle conditioned medium (MCM) or ordinary medium for 3 days, then the cells were collected for RNA, protein isolation and flow cytometry assay. ( a ) The mRNA expression of GRα and E1-C, E1-H was detected by RT-qPCR, n = 6 per group. ( b ) Expression of GRα protein was detected by Western blot (left) and displayed as column charts after quantification (right), n = 3 per group, * P

    Article Snippet: Antibodies against GRα (sc-393232, Santa Cruz, USA) and tubulin (sc-5274, Santa Cruz, USA) was used in Western blot analysis.

    Techniques: Expressing, Cell Culture, Isolation, Flow Cytometry, Cytometry, Quantitative RT-PCR, Western Blot

    GR levels in IMA were lower than SA. Backfat and longissimus dorsi muscles were from 12 Erhualian sows, then the adipose tissues were collected for RNA and protein isolation. ( a ) Copy numbers of total GR, GRα, GRβ mRNA in SA and IMA tissues were detected by RT-qPCR. ( b ) The ratio of GRα/GRβ was displayed according to the result obtained in ( a ). Data are shown as the mean ± SEM, n = 12 per group, * P

    Journal: Scientific Reports

    Article Title: Muscle-specific downregulation of GR levels inhibits adipogenesis in porcine intramuscular adipocyte tissue

    doi: 10.1038/s41598-017-00615-9

    Figure Lengend Snippet: GR levels in IMA were lower than SA. Backfat and longissimus dorsi muscles were from 12 Erhualian sows, then the adipose tissues were collected for RNA and protein isolation. ( a ) Copy numbers of total GR, GRα, GRβ mRNA in SA and IMA tissues were detected by RT-qPCR. ( b ) The ratio of GRα/GRβ was displayed according to the result obtained in ( a ). Data are shown as the mean ± SEM, n = 12 per group, * P

    Article Snippet: Antibodies against GRα (sc-393232, Santa Cruz, USA) and tubulin (sc-5274, Santa Cruz, USA) was used in Western blot analysis.

    Techniques: Isolation, Quantitative RT-PCR

    MSTN inhibited adipogenesis and reduced GRα expression. ( a ) The effects of MSTN on the proliferation of subcutaneous pre-adipocytes. Pre-adipocytes were treated with 50 ng/ml MSTN for 2 days, then the proliferating nuclei were stained red with EdU for 2 hours, and nuclei of all cells were stained blue with Hoechst (left). Three random pictures per group from confocal microscopy were used to count the cell numbers of EdU positive cells and Hoechst positive cells, and the ratio of the EdU positive cells to Hoechst positive cells was calculated in each picture (right). ( b ) Oil red O staining (left) and TG assay (right), n = 3 per group. The pre-adipocytes were cultured in culture medium until full confluence, then the cells were induced to differentiation with PBS or 50 ng/ml MSTN for 9 days. ( c ) The mRNA expression of GRα, exon 1C and 1H was detected by RT-qPCR, n = 6 per group. ( d ) The protein expression of GRα was detected by Western blot (left) and displayed as column charts after quantification (right), n = 3 per group. * P

    Journal: Scientific Reports

    Article Title: Muscle-specific downregulation of GR levels inhibits adipogenesis in porcine intramuscular adipocyte tissue

    doi: 10.1038/s41598-017-00615-9

    Figure Lengend Snippet: MSTN inhibited adipogenesis and reduced GRα expression. ( a ) The effects of MSTN on the proliferation of subcutaneous pre-adipocytes. Pre-adipocytes were treated with 50 ng/ml MSTN for 2 days, then the proliferating nuclei were stained red with EdU for 2 hours, and nuclei of all cells were stained blue with Hoechst (left). Three random pictures per group from confocal microscopy were used to count the cell numbers of EdU positive cells and Hoechst positive cells, and the ratio of the EdU positive cells to Hoechst positive cells was calculated in each picture (right). ( b ) Oil red O staining (left) and TG assay (right), n = 3 per group. The pre-adipocytes were cultured in culture medium until full confluence, then the cells were induced to differentiation with PBS or 50 ng/ml MSTN for 9 days. ( c ) The mRNA expression of GRα, exon 1C and 1H was detected by RT-qPCR, n = 6 per group. ( d ) The protein expression of GRα was detected by Western blot (left) and displayed as column charts after quantification (right), n = 3 per group. * P

    Article Snippet: Antibodies against GRα (sc-393232, Santa Cruz, USA) and tubulin (sc-5274, Santa Cruz, USA) was used in Western blot analysis.

    Techniques: Expressing, Staining, Confocal Microscopy, Cell Culture, Quantitative RT-PCR, Western Blot

    ISGs are expressed in the IECs of naive GR dim/dim mice. ( A – C ) RNA-seq of IECs of GR WT/WT (black) and GR dim/dim (white) mice ( n = 3 per group). ( A ) HOMER motif analysis of DE genes in GR dim/dim compared with GR WT/WT mice. Motifs with highest rank and their P values and q values are displayed. ( B ) Heat map of DE genes containing an ISRE and/or IRF-1 element. ( C ) Confirmation of RNA-seq data with qPCR on independent new samples ( n = 3 per group). Ifit1 , Irf8 , Irf1 , and Stat1 mRNA expression are shown as mean ± SEM. P values were calculated using Student’s t test. ( D – F ) GR WT/WT and GR dim/dim mice received drinking water with or without antibiotics for 3 weeks, after which IEC samples were taken ( n = 3 per group). ( D ) STAT1 and p-STAT1 protein levels were analyzed via Western blot using actin as a loading control. ( E ) Relative STAT1 and p-STAT1 signal intensities were quantified and normalized to ACTIN and STAT1 levels respectively. P values were calculated using 2-way ANOVA. ( F ) Stat1 and Ifit1 mRNA expression were determined in IECs of GR fl/fl (black) and GR VillinKO mice (white) via qPCR and are shown as mean ± SEM ( n = 5 per group). P values were calculated using Student’s t test. ( G ) GR recruitment to 2 IR-nGRE sites in the STAT1 promoter (IR-nGRE1 and IR-nGRE2). GR WT/WT (black) and GR dim/dim (white) mice were treated with PBS or 10 mg/kg DEX for 2 hours ( n = 5 per group; combined data of 3 independent experiments). ChIP on IEC samples was performed against GR using an H300 antibody. Data were normalized to input for each sample and expressed as fold change of H300 to IgG control. P values were calculated using 2-way ANOVA. **** P

    Journal: The Journal of Clinical Investigation

    Article Title: Glucocorticoid receptor dimers control intestinal STAT1 and TNF-induced inflammation in mice

    doi: 10.1172/JCI96636

    Figure Lengend Snippet: ISGs are expressed in the IECs of naive GR dim/dim mice. ( A – C ) RNA-seq of IECs of GR WT/WT (black) and GR dim/dim (white) mice ( n = 3 per group). ( A ) HOMER motif analysis of DE genes in GR dim/dim compared with GR WT/WT mice. Motifs with highest rank and their P values and q values are displayed. ( B ) Heat map of DE genes containing an ISRE and/or IRF-1 element. ( C ) Confirmation of RNA-seq data with qPCR on independent new samples ( n = 3 per group). Ifit1 , Irf8 , Irf1 , and Stat1 mRNA expression are shown as mean ± SEM. P values were calculated using Student’s t test. ( D – F ) GR WT/WT and GR dim/dim mice received drinking water with or without antibiotics for 3 weeks, after which IEC samples were taken ( n = 3 per group). ( D ) STAT1 and p-STAT1 protein levels were analyzed via Western blot using actin as a loading control. ( E ) Relative STAT1 and p-STAT1 signal intensities were quantified and normalized to ACTIN and STAT1 levels respectively. P values were calculated using 2-way ANOVA. ( F ) Stat1 and Ifit1 mRNA expression were determined in IECs of GR fl/fl (black) and GR VillinKO mice (white) via qPCR and are shown as mean ± SEM ( n = 5 per group). P values were calculated using Student’s t test. ( G ) GR recruitment to 2 IR-nGRE sites in the STAT1 promoter (IR-nGRE1 and IR-nGRE2). GR WT/WT (black) and GR dim/dim (white) mice were treated with PBS or 10 mg/kg DEX for 2 hours ( n = 5 per group; combined data of 3 independent experiments). ChIP on IEC samples was performed against GR using an H300 antibody. Data were normalized to input for each sample and expressed as fold change of H300 to IgG control. P values were calculated using 2-way ANOVA. **** P

    Article Snippet: Samples were first incubated with 5 μg anti-GR H300 antibody (catalog sc-8992; Santa Cruz Biotechnology Inc.) or IgG control (catalog 10500C; Invitrogen, Thermo Fisher Scientific) and then added to the blocked beads overnight.

    Techniques: Mouse Assay, RNA Sequencing Assay, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Chromatin Immunoprecipitation

    Dex increases the abundance of nuclear GR. Western blot analysis using a polyclonal anti-GR antibody were performed on nuclear extracts of A549 cells. ( A ) Cells were exposed to vehicle (−) or 100 nM Dex (+) for 24 h; 7 or 14 μg of nuclear protein (NP) were analyzed. ( B ) Cells were exposed to 100 nM Dex for the times indicated. Ten micrograms of NP were evaluated for GR and actin protein for each time point. Results are shown as the relative fold induction of Dex-induced nuclear GR levels normalized to actin levels, expressed as the mean ± SE. Experiments were performed in triplicate on three separate occasions. * P

    Journal: American Journal of Respiratory Cell and Molecular Biology

    Article Title: Dexamethasone-Mediated Repression of MUC5AC Gene Expression in Human Lung Epithelial Cells

    doi: 10.1165/rcmb.2005-0176OC

    Figure Lengend Snippet: Dex increases the abundance of nuclear GR. Western blot analysis using a polyclonal anti-GR antibody were performed on nuclear extracts of A549 cells. ( A ) Cells were exposed to vehicle (−) or 100 nM Dex (+) for 24 h; 7 or 14 μg of nuclear protein (NP) were analyzed. ( B ) Cells were exposed to 100 nM Dex for the times indicated. Ten micrograms of NP were evaluated for GR and actin protein for each time point. Results are shown as the relative fold induction of Dex-induced nuclear GR levels normalized to actin levels, expressed as the mean ± SE. Experiments were performed in triplicate on three separate occasions. * P

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and immunoblotted with polyclonal anti-GR (1:1,000) or monoclonal anti–actin (1:1,000) antibody (Santa Cruz Biotechnology, Santa Cruz, CA).

    Techniques: Western Blot