nurr1 Search Results


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  • 91
    Thermo Fisher mouse monoclonal antibody against nurr 1
    Effect of Nor-1, <t>Nurr-1</t> and Nur-77 silencing on forskolin-mediated BeWo cell fusion and hCG secretion. BeWo cell knockdown for Nor-1, Nurr-1 or Nur-77 were made using siRNA as described in the Methods section. The efficacy of silencing of Nor-1, Nurr-1 or Nur-77 transcripts was confirmed via qRT-PCR using specific primers and was studied after 0, 2 and 48 h of forskolin (25 μM) treatment in control siRNA-transfected and Nor-1-, Nurr-1- or Nur-77-silenced BeWo cells. a , b and c – Comparisons of the transcript levels of Nor-1, Nurr-1 and Nur-77, respectively, in the control siRNA-transfected and silenced cells. Each bar represents relative expression after normalization with 18S rRNA and is expressed as means ± s.e.m. of three independent experiments performed in triplicate. The effect of Nor-1, Nurr-1 or Nur-77 silencing on forskolin-mediated BeWo cell fusion was studied at 48 h using desmoplakin I + II staining and hCG secretion was studied with ELISA. d – The fold change in forskolin-mediated BeWo cell fusion in Nor-1-, Nurr-1- or Nur-77-silenced cells and control siRNA-transfected cells. Values are shown as the means ± s.e.m. of three independent experiments. Representative images of desmoplakin I + II staining (green) are appended alongside. The scale bar represents 20 μm. e – hCG secreted by control and Nor-1-, Nurr-1- or Nur-77-silenced cells in response to forskolin treatment at 48 h and represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant
    Mouse Monoclonal Antibody Against Nurr 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore anti nurr1
    Activation of <t>Nurr1</t> with compounds 1 and 2 . (A) Luciferase assay to measure Nurr1 activity with 1 , (B) with 2 . Experiments were performed with human neuroblastoma SK-N-BE(2)C cells transfected with pTH2600LUC, pCMV Nurr1-myc, pZeoSV-Nurr1(LBD), and p8xGAL-Luc for the luciferase assay. (C) Luciferase assay to measure Nurr1 activity with 1 and 2 at 0.3 μ M in Nurr1-knockdown cells. Human neuroblastoma SK-N-BE(2)C cells were transfected with pTH2600LUC, pCMV Nurr1-myc, pZeoSV-Nurr1(LBD), p8xGAL-Luc, and Nurr1-siRNA for the luciferase assay. (D) Western blotting analysis of Nurr1 in Nurr1-siRNA- treated SK-N-BE(2)C cells.
    Anti Nurr1, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology nurr 1
    Effects of gene transfer on striatal dopaminergic innervation. The column on the left illustrates sections throughout the striatum of treated animals immunoreacted with an antibody raised against TH, to reveal the density of TH-immunopositive fibers in the striatum. The lesioned side is on the left, the control side, on the right. Notice that only animals injected with RAd-GDNF showed a protection of the striatal dopaminergic fibers. The graph shows the densitometric analysis of TH + fiber density in the striatum of rats treated with 1 × 10 8 IU of RAd-Shh, RAd-Gli-1, RAd-GDNF, or RAd-35. Degeneration of axonal terminals in the striatum after administration of 6-OHDA is not prevented following the injection of RAd-ShhN, RAd-Gli-1, or <t>RAd-Nurr-1</t> (not shown). Only RAd-GDNF is able to protect TH + fibers significantly. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05. Scale bar, 1 mm.
    Nurr 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Perseus Proteomics nurr 1
    Effects of gene transfer on striatal dopaminergic innervation. The column on the left illustrates sections throughout the striatum of treated animals immunoreacted with an antibody raised against TH, to reveal the density of TH-immunopositive fibers in the striatum. The lesioned side is on the left, the control side, on the right. Notice that only animals injected with RAd-GDNF showed a protection of the striatal dopaminergic fibers. The graph shows the densitometric analysis of TH + fiber density in the striatum of rats treated with 1 × 10 8 IU of RAd-Shh, RAd-Gli-1, RAd-GDNF, or RAd-35. Degeneration of axonal terminals in the striatum after administration of 6-OHDA is not prevented following the injection of RAd-ShhN, RAd-Gli-1, or <t>RAd-Nurr-1</t> (not shown). Only RAd-GDNF is able to protect TH + fibers significantly. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05. Scale bar, 1 mm.
    Nurr 1, supplied by Perseus Proteomics, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Santa Cruz Biotechnology anti nurr 1 antibody
    Effects of gene transfer on striatal dopaminergic innervation. The column on the left illustrates sections throughout the striatum of treated animals immunoreacted with an antibody raised against TH, to reveal the density of TH-immunopositive fibers in the striatum. The lesioned side is on the left, the control side, on the right. Notice that only animals injected with RAd-GDNF showed a protection of the striatal dopaminergic fibers. The graph shows the densitometric analysis of TH + fiber density in the striatum of rats treated with 1 × 10 8 IU of RAd-Shh, RAd-Gli-1, RAd-GDNF, or RAd-35. Degeneration of axonal terminals in the striatum after administration of 6-OHDA is not prevented following the injection of RAd-ShhN, RAd-Gli-1, or <t>RAd-Nurr-1</t> (not shown). Only RAd-GDNF is able to protect TH + fibers significantly. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05. Scale bar, 1 mm.
    Anti Nurr 1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam anti nurr 1
    Effects of gene transfer on striatal dopaminergic innervation. The column on the left illustrates sections throughout the striatum of treated animals immunoreacted with an antibody raised against TH, to reveal the density of TH-immunopositive fibers in the striatum. The lesioned side is on the left, the control side, on the right. Notice that only animals injected with RAd-GDNF showed a protection of the striatal dopaminergic fibers. The graph shows the densitometric analysis of TH + fiber density in the striatum of rats treated with 1 × 10 8 IU of RAd-Shh, RAd-Gli-1, RAd-GDNF, or RAd-35. Degeneration of axonal terminals in the striatum after administration of 6-OHDA is not prevented following the injection of RAd-ShhN, RAd-Gli-1, or <t>RAd-Nurr-1</t> (not shown). Only RAd-GDNF is able to protect TH + fibers significantly. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05. Scale bar, 1 mm.
    Anti Nurr 1, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam antibody nurr1
    The impact of miR-145 overexpression and <t>Nurr1</t> on the differentiation of NSCs induced by BMP2. NSCs were treated with 0 ng/ml of BMP2, or 20 ng/ml of BMP2, or 20 ng/ml of BMP2+NC, or 20 ng/ml of BMP2+miR-145 mimics, or 20 ng/ml of BMP2+miR-145 mimics+pcDNA-3.1,
    Antibody Nurr1, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology nr4a2 nurr 1 antibody
    The impact of miR-145 overexpression and <t>Nurr1</t> on the differentiation of NSCs induced by BMP2. NSCs were treated with 0 ng/ml of BMP2, or 20 ng/ml of BMP2, or 20 ng/ml of BMP2+NC, or 20 ng/ml of BMP2+miR-145 mimics, or 20 ng/ml of BMP2+miR-145 mimics+pcDNA-3.1,
    Nr4a2 Nurr 1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore nurr1
    Exogenous expression of A53T α-syn suppresses the nuclear accumulation of <t>Nurr1</t> and the expression of Nurr1-controlled dopaminergic genes ( A ) Nurr1 (green) and TH (red) co-staining in the SNC coronal sections of 1-month-old of nTg , A53T , and DOX-treated A53T ( A53T/DOX ) mice. Asterisks indicate TH-positive neurons with severe loss of Nurr1 staining. Arrows mark the mDA neurons with intensified Nurr1 staining. Arrowheads point to non-DA cells stained with Nurr1. Scale bar: 20 μm ( B ) The intensity of Nurr1 signals in the nucleus of SNC mDA neurons from 1-month-old nTg , A53T , and A53T/DOX mice (n ≥ 3 animals per genotype; ≥ 40 neurons per animal). Data were presented as Mean ± SEM. ***p
    Nurr1, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology nurr1 nr4a
    Exogenous expression of A53T α-syn suppresses the nuclear accumulation of <t>Nurr1</t> and the expression of Nurr1-controlled dopaminergic genes ( A ) Nurr1 (green) and TH (red) co-staining in the SNC coronal sections of 1-month-old of nTg , A53T , and DOX-treated A53T ( A53T/DOX ) mice. Asterisks indicate TH-positive neurons with severe loss of Nurr1 staining. Arrows mark the mDA neurons with intensified Nurr1 staining. Arrowheads point to non-DA cells stained with Nurr1. Scale bar: 20 μm ( B ) The intensity of Nurr1 signals in the nucleus of SNC mDA neurons from 1-month-old nTg , A53T , and A53T/DOX mice (n ≥ 3 animals per genotype; ≥ 40 neurons per animal). Data were presented as Mean ± SEM. ***p
    Nurr1 Nr4a, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Santa Cruz Biotechnology α nurr1
    Exogenous expression of A53T α-syn suppresses the nuclear accumulation of <t>Nurr1</t> and the expression of Nurr1-controlled dopaminergic genes ( A ) Nurr1 (green) and TH (red) co-staining in the SNC coronal sections of 1-month-old of nTg , A53T , and DOX-treated A53T ( A53T/DOX ) mice. Asterisks indicate TH-positive neurons with severe loss of Nurr1 staining. Arrows mark the mDA neurons with intensified Nurr1 staining. Arrowheads point to non-DA cells stained with Nurr1. Scale bar: 20 μm ( B ) The intensity of Nurr1 signals in the nucleus of SNC mDA neurons from 1-month-old nTg , A53T , and A53T/DOX mice (n ≥ 3 animals per genotype; ≥ 40 neurons per animal). Data were presented as Mean ± SEM. ***p
    α Nurr1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher nurr1
    METH caused differential changes in <t>Nurr1</t> and Pitx3 mRNA and protein levels in the midbrain. METH injections resulted in significant decreases in Nurr1 expression in all the groups (A). In contrast, METH challenges caused significant increases in Pitx3 mRNA expression only in the METH-preconditioned rats (B). Consistent with the mRNA data, toxic METH challenges caused decreases in Nurr1 protein levels in all the groups (C). In contrast, Pitx3 protein levels were increased by METH (D). The values represent means ± SEM in comparison to the saline-pretreated challenged with saline group (SSS). N = 3–6 animals per group. Keys to statistics are as described in the legend to Fig. 4 .
    Nurr1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher anti nurr1
    Protein expression of synapsins, CREB, and <t>Nurr1</t> in mouse primary cortical neurons following IC delivery of recombinant h-αSyn WT species. ( A ) IC delivery of exogenous fluorophore-conjugated antibodies using Chariot in primary cortical neurons 60 min postapplication. Neurons were labeled with MAP2 (magenta). (Scale bars: 10 μm.) ( B ) Representative Western blot (WB) images for αSyn in selected fractions following SEC segregation of recombinant monomers and multimers. Fraction #50 was enriched in monomers, whereas #42 was enriched in αSyn multimers. ( C ) Representative Western blot images for αSyn using lysates of cells subjected to IC delivery of fractions #50 and #42 (250 nM, monomer equivalent). ( D ) Representative Western blot analysis of synapsin expression in primary neurons 6 h after intraneuronal delivery of recombinant monomers or oligomers isolated by SEC. Actin was used as internal loading control. ( E ) Quantification of the relative expression levels of synapsins following intraneuronal delivery of recombinant h-αSyn WT species. (ANOVA followed by Student t test with Bonferroni correction, n = 4–6 dishes per group.) ( F ) Western blot analysis with 4D6 antibodies following Clear Native (CN)-PAGE segregation of SEC fraction #48 using IC lysates from TgI2.2, WT, or SNCA-null (KO) mice. Oligomeric recombinant h-αSyn WT (0.25 μg) was used as control. ( G ) Representative Western blot images documenting the protein abundance for pS133-CREB, total CREB, Nurr1, and the neuronal nuclear protein NeuN in membrane lysates from primary neurons treated with αSyn species delivered with Chariot. Please note the selective reductions in phosphorylated (p)CREB and Nurr1, whereas NeuN amounts remain unchanged. Veh., vehicle.
    Anti Nurr1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nurr1  (Abcam)
    91
    Abcam nurr1
    Effects of intrastriatal injection of <t>Lv-Nurr1-MSCs</t> on apomorphine-induced rotation asymmetry in rats after 6-hydroxydopamine (6-OHDA) lesion. Average rotational turns/min are showed at different time points (1, 2 and 4 weeks, n = 5 for each group). The average rotational rates of PD rats in the Lv-MSCs group (10.57 ± 1.86 at 1 week, 7.64 ± 1.56 at 2 weeks and 6.71 ± 1.87 at 4 weeks) significantly decreased ( # P
    Nurr1, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of Nor-1, Nurr-1 and Nur-77 silencing on forskolin-mediated BeWo cell fusion and hCG secretion. BeWo cell knockdown for Nor-1, Nurr-1 or Nur-77 were made using siRNA as described in the Methods section. The efficacy of silencing of Nor-1, Nurr-1 or Nur-77 transcripts was confirmed via qRT-PCR using specific primers and was studied after 0, 2 and 48 h of forskolin (25 μM) treatment in control siRNA-transfected and Nor-1-, Nurr-1- or Nur-77-silenced BeWo cells. a , b and c – Comparisons of the transcript levels of Nor-1, Nurr-1 and Nur-77, respectively, in the control siRNA-transfected and silenced cells. Each bar represents relative expression after normalization with 18S rRNA and is expressed as means ± s.e.m. of three independent experiments performed in triplicate. The effect of Nor-1, Nurr-1 or Nur-77 silencing on forskolin-mediated BeWo cell fusion was studied at 48 h using desmoplakin I + II staining and hCG secretion was studied with ELISA. d – The fold change in forskolin-mediated BeWo cell fusion in Nor-1-, Nurr-1- or Nur-77-silenced cells and control siRNA-transfected cells. Values are shown as the means ± s.e.m. of three independent experiments. Representative images of desmoplakin I + II staining (green) are appended alongside. The scale bar represents 20 μm. e – hCG secreted by control and Nor-1-, Nurr-1- or Nur-77-silenced cells in response to forskolin treatment at 48 h and represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant

    Journal: Cellular & Molecular Biology Letters

    Article Title: Relevance of the NR4A sub-family of nuclear orphan receptors in trophoblastic BeWo cell differentiation

    doi: 10.1186/s11658-017-0046-0

    Figure Lengend Snippet: Effect of Nor-1, Nurr-1 and Nur-77 silencing on forskolin-mediated BeWo cell fusion and hCG secretion. BeWo cell knockdown for Nor-1, Nurr-1 or Nur-77 were made using siRNA as described in the Methods section. The efficacy of silencing of Nor-1, Nurr-1 or Nur-77 transcripts was confirmed via qRT-PCR using specific primers and was studied after 0, 2 and 48 h of forskolin (25 μM) treatment in control siRNA-transfected and Nor-1-, Nurr-1- or Nur-77-silenced BeWo cells. a , b and c – Comparisons of the transcript levels of Nor-1, Nurr-1 and Nur-77, respectively, in the control siRNA-transfected and silenced cells. Each bar represents relative expression after normalization with 18S rRNA and is expressed as means ± s.e.m. of three independent experiments performed in triplicate. The effect of Nor-1, Nurr-1 or Nur-77 silencing on forskolin-mediated BeWo cell fusion was studied at 48 h using desmoplakin I + II staining and hCG secretion was studied with ELISA. d – The fold change in forskolin-mediated BeWo cell fusion in Nor-1-, Nurr-1- or Nur-77-silenced cells and control siRNA-transfected cells. Values are shown as the means ± s.e.m. of three independent experiments. Representative images of desmoplakin I + II staining (green) are appended alongside. The scale bar represents 20 μm. e – hCG secreted by control and Nor-1-, Nurr-1- or Nur-77-silenced cells in response to forskolin treatment at 48 h and represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant

    Article Snippet: After protein transfer, the nitrocellulose membrane was blocked in 5% BSA in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20; pH -7.4) and further incubated at 4 °C overnight with an optimized dilution of 1:1000 of rabbit polyclonal antibodies against Nor-1, Nur-77 (Thermo Fisher Scientific) and GAPDH (Cell Signaling Technology Inc.), and mouse monoclonal antibody against Nurr-1 (Thermo Fisher Scientific) in TBST containing 2.5% BSA.

    Techniques: Quantitative RT-PCR, Transfection, Expressing, Staining, Enzyme-linked Immunosorbent Assay

    Transcript profiles of NR4A members in BeWo cells transfected with Nor-1, Nurr-1 or Nur-77 siRNA after forskolin treatment. BeWo cells were silenced for Nor-1, Nurr-1 or Nur-77 using the respective siRNA and transcript levels of all the three members were assessed using qRT-PCR after 0, 2 and 48 h of forskolin (25 μM) treatment. a , b and c – qRT-PCR analysis of the transcript levels of all three members in BeWo cells with Nor-1, Nurr-1 and Nur-77 knockdown, respectively. Each bar represents the relative expression after normalization with 18S rRNA, expressed as means ± s.e.m. of three independent experiments performed in triplicate. p ≤ 0.05 is considered statistically significant

    Journal: Cellular & Molecular Biology Letters

    Article Title: Relevance of the NR4A sub-family of nuclear orphan receptors in trophoblastic BeWo cell differentiation

    doi: 10.1186/s11658-017-0046-0

    Figure Lengend Snippet: Transcript profiles of NR4A members in BeWo cells transfected with Nor-1, Nurr-1 or Nur-77 siRNA after forskolin treatment. BeWo cells were silenced for Nor-1, Nurr-1 or Nur-77 using the respective siRNA and transcript levels of all the three members were assessed using qRT-PCR after 0, 2 and 48 h of forskolin (25 μM) treatment. a , b and c – qRT-PCR analysis of the transcript levels of all three members in BeWo cells with Nor-1, Nurr-1 and Nur-77 knockdown, respectively. Each bar represents the relative expression after normalization with 18S rRNA, expressed as means ± s.e.m. of three independent experiments performed in triplicate. p ≤ 0.05 is considered statistically significant

    Article Snippet: After protein transfer, the nitrocellulose membrane was blocked in 5% BSA in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20; pH -7.4) and further incubated at 4 °C overnight with an optimized dilution of 1:1000 of rabbit polyclonal antibodies against Nor-1, Nur-77 (Thermo Fisher Scientific) and GAPDH (Cell Signaling Technology Inc.), and mouse monoclonal antibody against Nurr-1 (Thermo Fisher Scientific) in TBST containing 2.5% BSA.

    Techniques: Transfection, Quantitative RT-PCR, Expressing

    Effect of Nur-77 knockdown on hCG- and GnRH-mediated cell fusion and hCG secretion. BeWo cells were treated with either hCG (5 IU/ml) or GnRH (10 ng) for various times. Subsequently, total RNA and cell lysates were isolated and processed for qRT-PCR and Western blotting, respectively, to analyze the transcript and protein levels of Nor-1, Nurr-1 and Nur-77. a and b – qRT-PCR analysis of Nor-1, Nurr-1 and Nur-77 in the form of bar graphs of BeWo cells treated with hCG and GnRH, respectively. Relative expression was normalized with 18S rRNA. Data are expressed as means ± s.e.m. of three independent experiments performed in triplicate. c and d – Protein expression profiles of Nor-1, Nurr-1 and Nur-77 in BeWo cells after treatment with hCG and GnRH, respectively, with GAPDH used as an internal control. Values are expressed as means ± s.e.m. of the band intensity of three independent experiments. Representative blots for the same experiment are appended with the graphs. BeWo cells transfected with either Nur-77 siRNA or control siRNA were treated with either hCG (5 IU/ml) or GnRH (10 ng/ml). The fold change in fusion was estimated after 48 h using desmoplakin I + II staining and hCG secretion was analyzed in GnRH treated cells via ELISA. e – The effect of hCG and GnRH treatment in the control and Nur-77-silenced BeWo cells on fusion at 48 h. The data are expressed as means ± s.e.m. of three independent experiments. Representative images of desmoplakin I + II staining in green and DAPI in blue are appended alongside. The scale bar is 20 μm. f – hCG secreted by control and Nur-77-silenced BeWo cells in response to GnRH treatment at 48 h. Data are represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant

    Journal: Cellular & Molecular Biology Letters

    Article Title: Relevance of the NR4A sub-family of nuclear orphan receptors in trophoblastic BeWo cell differentiation

    doi: 10.1186/s11658-017-0046-0

    Figure Lengend Snippet: Effect of Nur-77 knockdown on hCG- and GnRH-mediated cell fusion and hCG secretion. BeWo cells were treated with either hCG (5 IU/ml) or GnRH (10 ng) for various times. Subsequently, total RNA and cell lysates were isolated and processed for qRT-PCR and Western blotting, respectively, to analyze the transcript and protein levels of Nor-1, Nurr-1 and Nur-77. a and b – qRT-PCR analysis of Nor-1, Nurr-1 and Nur-77 in the form of bar graphs of BeWo cells treated with hCG and GnRH, respectively. Relative expression was normalized with 18S rRNA. Data are expressed as means ± s.e.m. of three independent experiments performed in triplicate. c and d – Protein expression profiles of Nor-1, Nurr-1 and Nur-77 in BeWo cells after treatment with hCG and GnRH, respectively, with GAPDH used as an internal control. Values are expressed as means ± s.e.m. of the band intensity of three independent experiments. Representative blots for the same experiment are appended with the graphs. BeWo cells transfected with either Nur-77 siRNA or control siRNA were treated with either hCG (5 IU/ml) or GnRH (10 ng/ml). The fold change in fusion was estimated after 48 h using desmoplakin I + II staining and hCG secretion was analyzed in GnRH treated cells via ELISA. e – The effect of hCG and GnRH treatment in the control and Nur-77-silenced BeWo cells on fusion at 48 h. The data are expressed as means ± s.e.m. of three independent experiments. Representative images of desmoplakin I + II staining in green and DAPI in blue are appended alongside. The scale bar is 20 μm. f – hCG secreted by control and Nur-77-silenced BeWo cells in response to GnRH treatment at 48 h. Data are represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant

    Article Snippet: After protein transfer, the nitrocellulose membrane was blocked in 5% BSA in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20; pH -7.4) and further incubated at 4 °C overnight with an optimized dilution of 1:1000 of rabbit polyclonal antibodies against Nor-1, Nur-77 (Thermo Fisher Scientific) and GAPDH (Cell Signaling Technology Inc.), and mouse monoclonal antibody against Nurr-1 (Thermo Fisher Scientific) in TBST containing 2.5% BSA.

    Techniques: Isolation, Quantitative RT-PCR, Western Blot, Expressing, Transfection, Staining, Enzyme-linked Immunosorbent Assay

    Expression profile of the members of the NR4A sub-family in BeWo cells treated with forskolin. BeWo cells were treated with forskolin (25 μM) for various time periods, followed by analysis of NR4A sub-family members at the mRNA and protein levels. a – Transcript profiles of Nor-1, Nurr-1 and Nur-77 in BeWo cells at 0, 1, 2, 6, 24 and 48 h of forskolin treatment. Relative expression was normalized with 18S rRNA. Data are expressed as means ± s.e.m. of three independent experiments performed in triplicate. b – Densitometeric plots of Nor-1, Nurr-1 and Nur-77 using GAPDH as a loading control. Data are represented as means ± s.e.m. of at least three experiments. Representative blots for the same are also shown. p ≤ 0.05 is considered statistically significant

    Journal: Cellular & Molecular Biology Letters

    Article Title: Relevance of the NR4A sub-family of nuclear orphan receptors in trophoblastic BeWo cell differentiation

    doi: 10.1186/s11658-017-0046-0

    Figure Lengend Snippet: Expression profile of the members of the NR4A sub-family in BeWo cells treated with forskolin. BeWo cells were treated with forskolin (25 μM) for various time periods, followed by analysis of NR4A sub-family members at the mRNA and protein levels. a – Transcript profiles of Nor-1, Nurr-1 and Nur-77 in BeWo cells at 0, 1, 2, 6, 24 and 48 h of forskolin treatment. Relative expression was normalized with 18S rRNA. Data are expressed as means ± s.e.m. of three independent experiments performed in triplicate. b – Densitometeric plots of Nor-1, Nurr-1 and Nur-77 using GAPDH as a loading control. Data are represented as means ± s.e.m. of at least three experiments. Representative blots for the same are also shown. p ≤ 0.05 is considered statistically significant

    Article Snippet: After protein transfer, the nitrocellulose membrane was blocked in 5% BSA in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20; pH -7.4) and further incubated at 4 °C overnight with an optimized dilution of 1:1000 of rabbit polyclonal antibodies against Nor-1, Nur-77 (Thermo Fisher Scientific) and GAPDH (Cell Signaling Technology Inc.), and mouse monoclonal antibody against Nurr-1 (Thermo Fisher Scientific) in TBST containing 2.5% BSA.

    Techniques: Expressing

    Effect of simultaneous silencing of Nor-1, Nurr-1 and Nur-77 on forskolin- or hCG-mediated BeWo cell differentiation. BeWo cells were knocked down for Nor-1, Nurr-1 and Nur-77 simultaneously using siRNA as described in Methods section. To confirm target-specific silencing of all three nuclear orphan receptors, qRT-PCR was performed at 0, 2 and 48 h after forskolin treatment in control siRNA-transfected and silenced cells. The effect of NR4A silencing on BeWo cell fusion and/or hCG secretion was studied after 48 h of forskolin and hCG treatment. a – Comparison of the transcript levels of Nor-1, Nurr-1 and Nur-77 in the control siRNA-transfected and NR4A-silenced cells. Each bar represents relative expression after normalization with 18S rRNA, expressed as means ± s.e.m. of three independent experiments performed in triplicate. b – The fold change in forskolin-mediated BeWo cell fusion in control siRNA-transfected cells and NR4A-silenced cells. Values are shown as means ± s.e.m. of three independent experiments. c – hCG secreted by control and NR4A-silenced cells on treatment with forskolin for 48 h. Data are represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant

    Journal: Cellular & Molecular Biology Letters

    Article Title: Relevance of the NR4A sub-family of nuclear orphan receptors in trophoblastic BeWo cell differentiation

    doi: 10.1186/s11658-017-0046-0

    Figure Lengend Snippet: Effect of simultaneous silencing of Nor-1, Nurr-1 and Nur-77 on forskolin- or hCG-mediated BeWo cell differentiation. BeWo cells were knocked down for Nor-1, Nurr-1 and Nur-77 simultaneously using siRNA as described in Methods section. To confirm target-specific silencing of all three nuclear orphan receptors, qRT-PCR was performed at 0, 2 and 48 h after forskolin treatment in control siRNA-transfected and silenced cells. The effect of NR4A silencing on BeWo cell fusion and/or hCG secretion was studied after 48 h of forskolin and hCG treatment. a – Comparison of the transcript levels of Nor-1, Nurr-1 and Nur-77 in the control siRNA-transfected and NR4A-silenced cells. Each bar represents relative expression after normalization with 18S rRNA, expressed as means ± s.e.m. of three independent experiments performed in triplicate. b – The fold change in forskolin-mediated BeWo cell fusion in control siRNA-transfected cells and NR4A-silenced cells. Values are shown as means ± s.e.m. of three independent experiments. c – hCG secreted by control and NR4A-silenced cells on treatment with forskolin for 48 h. Data are represented as means ± s.e.m. of three independent experiments performed in duplicate. p ≤ 0.05 is considered statistically significant

    Article Snippet: After protein transfer, the nitrocellulose membrane was blocked in 5% BSA in TBST (50 mM Tris-HCl, 150 mM NaCl, 0.1% Tween-20; pH -7.4) and further incubated at 4 °C overnight with an optimized dilution of 1:1000 of rabbit polyclonal antibodies against Nor-1, Nur-77 (Thermo Fisher Scientific) and GAPDH (Cell Signaling Technology Inc.), and mouse monoclonal antibody against Nurr-1 (Thermo Fisher Scientific) in TBST containing 2.5% BSA.

    Techniques: Cell Differentiation, Quantitative RT-PCR, Transfection, Expressing

    Activation of Nurr1 with compounds 1 and 2 . (A) Luciferase assay to measure Nurr1 activity with 1 , (B) with 2 . Experiments were performed with human neuroblastoma SK-N-BE(2)C cells transfected with pTH2600LUC, pCMV Nurr1-myc, pZeoSV-Nurr1(LBD), and p8xGAL-Luc for the luciferase assay. (C) Luciferase assay to measure Nurr1 activity with 1 and 2 at 0.3 μ M in Nurr1-knockdown cells. Human neuroblastoma SK-N-BE(2)C cells were transfected with pTH2600LUC, pCMV Nurr1-myc, pZeoSV-Nurr1(LBD), p8xGAL-Luc, and Nurr1-siRNA for the luciferase assay. (D) Western blotting analysis of Nurr1 in Nurr1-siRNA- treated SK-N-BE(2)C cells.

    Journal: Journal of natural products

    Article Title: Daphnane Diterpenes from Daphne genkwa Activate Nurr1 and Have a Neuroprotective Effect in an Animal Model of Parkinson’s Disease

    doi: 10.1021/acs.jnatprod.6b00110

    Figure Lengend Snippet: Activation of Nurr1 with compounds 1 and 2 . (A) Luciferase assay to measure Nurr1 activity with 1 , (B) with 2 . Experiments were performed with human neuroblastoma SK-N-BE(2)C cells transfected with pTH2600LUC, pCMV Nurr1-myc, pZeoSV-Nurr1(LBD), and p8xGAL-Luc for the luciferase assay. (C) Luciferase assay to measure Nurr1 activity with 1 and 2 at 0.3 μ M in Nurr1-knockdown cells. Human neuroblastoma SK-N-BE(2)C cells were transfected with pTH2600LUC, pCMV Nurr1-myc, pZeoSV-Nurr1(LBD), p8xGAL-Luc, and Nurr1-siRNA for the luciferase assay. (D) Western blotting analysis of Nurr1 in Nurr1-siRNA- treated SK-N-BE(2)C cells.

    Article Snippet: The following primary antibodies were used: rabbit anticleaved caspase-3 [Cell Signaling (Danver, MA, U.S.A.); 1:1000], anticytochrome c [Pharmingen (San Diego, CA, U.S.A.)], anti-interleukin 1 β [Santa Cruz (Santa Cruz, CA, U.S.A.)], anti-nurr1 (Sigma), anti-SOD-1 (Santa Cruz), and mouse antiactin (Sigma 1:5000).

    Techniques: Activation Assay, Luciferase, Activity Assay, Transfection, Western Blot

    Nur77 and Nurr1 form heterodimers. (A) The abilities of Nur77 and Nurr1 to interact in vitro were assessed by using a pull-down assay. Fusion proteins consisting of MBP and either Nur77 or LacZ (as a control) were bound to a maltose column, and either in vitro-translated Nurr1 or luciferase (Luc; as a control) was tested for binding. The position of full-length Nurr1 (61 kDa) is indicated by an arrow. (B) Formation of Nurr1-Nur77 heterodimer complexes upon binding of a NurRE CON probe in a gel retardation assay. Since native proteins comigrated in gel retardation experiments (data not shown), we used a mutant Nur77 with a truncated C-terminal domain for the experiment; this mutant preferentially binds as a monomer on its own (lanes 1 to 3), whereas Nurr1 binds both as a monomer and as a homodimer (lane 4). In the presence of increasing amounts of mutant Nur77 (lanes 5 to 7) and Nurr1, a new complex (labeled heterodimer) appears (lane 7). Formation of these heterodimers is blocked by an antiserum specific for Nurr1 (lane 10) or Nur77 (lane 11). The anti-Nurr1 does not recognize Nur77 (lane 8), and the anti-Nur77 does not recognize Nurr1 (lane 9). (C and D) Two-hybrid assays were performed in CV1 cells to show that the chimeric proteins Gal4-Nur77 and VP16-Nurr1 (C) or Gal4-Nurr1 and VP16-Nur77 (D) interact in vivo. Cells were cotransfected with a UAS-containing luciferase reporter plasmid. (E) Presence of Nur77-Nurr1 complexes, as revealed by coimmunoprecipitation of AtT-20 cell nuclear extracts. Extracts were immunoprecipitated with antibodies against Nur77 (lane 1) or control IgG (lane 2) and analyzed by Western blotting with Nurr1 antiserum. In vitro-translated Nurr1 protein was used as a reference (lane 3).

    Journal: Molecular and Cellular Biology

    Article Title: Heterodimerization between Members of the Nur Subfamily of Orphan Nuclear Receptors as a Novel Mechanism for Gene Activation

    doi:

    Figure Lengend Snippet: Nur77 and Nurr1 form heterodimers. (A) The abilities of Nur77 and Nurr1 to interact in vitro were assessed by using a pull-down assay. Fusion proteins consisting of MBP and either Nur77 or LacZ (as a control) were bound to a maltose column, and either in vitro-translated Nurr1 or luciferase (Luc; as a control) was tested for binding. The position of full-length Nurr1 (61 kDa) is indicated by an arrow. (B) Formation of Nurr1-Nur77 heterodimer complexes upon binding of a NurRE CON probe in a gel retardation assay. Since native proteins comigrated in gel retardation experiments (data not shown), we used a mutant Nur77 with a truncated C-terminal domain for the experiment; this mutant preferentially binds as a monomer on its own (lanes 1 to 3), whereas Nurr1 binds both as a monomer and as a homodimer (lane 4). In the presence of increasing amounts of mutant Nur77 (lanes 5 to 7) and Nurr1, a new complex (labeled heterodimer) appears (lane 7). Formation of these heterodimers is blocked by an antiserum specific for Nurr1 (lane 10) or Nur77 (lane 11). The anti-Nurr1 does not recognize Nur77 (lane 8), and the anti-Nur77 does not recognize Nurr1 (lane 9). (C and D) Two-hybrid assays were performed in CV1 cells to show that the chimeric proteins Gal4-Nur77 and VP16-Nurr1 (C) or Gal4-Nurr1 and VP16-Nur77 (D) interact in vivo. Cells were cotransfected with a UAS-containing luciferase reporter plasmid. (E) Presence of Nur77-Nurr1 complexes, as revealed by coimmunoprecipitation of AtT-20 cell nuclear extracts. Extracts were immunoprecipitated with antibodies against Nur77 (lane 1) or control IgG (lane 2) and analyzed by Western blotting with Nurr1 antiserum. In vitro-translated Nurr1 protein was used as a reference (lane 3).

    Article Snippet: Nurr1 was revealed by Western blotting with a Nurr1 antibody (a gift from T. Perlmann) and an anti-rabbit antibody–horseradish peroxidase conjugate (Sigma).

    Techniques: In Vitro, Pull Down Assay, Luciferase, Binding Assay, Electrophoretic Mobility Shift Assay, Mutagenesis, Labeling, In Vivo, Plasmid Preparation, Immunoprecipitation, Western Blot

    Induction of Nur77, Nurr1, and NOR-1 mRNAs by CRH in pituitary-derived AtT-20 cells. The effect of CRH (10 −7 M), alone or in combination with DEX (10 −7 M), on Nur77, Nurr1, and NOR-1 mRNAs was measured by Northern blotting. RNA was extracted from cells treated or untreated (Ctl) for the indicated time periods. β-Actin mRNA was used as a loading control.

    Journal: Molecular and Cellular Biology

    Article Title: Heterodimerization between Members of the Nur Subfamily of Orphan Nuclear Receptors as a Novel Mechanism for Gene Activation

    doi:

    Figure Lengend Snippet: Induction of Nur77, Nurr1, and NOR-1 mRNAs by CRH in pituitary-derived AtT-20 cells. The effect of CRH (10 −7 M), alone or in combination with DEX (10 −7 M), on Nur77, Nurr1, and NOR-1 mRNAs was measured by Northern blotting. RNA was extracted from cells treated or untreated (Ctl) for the indicated time periods. β-Actin mRNA was used as a loading control.

    Article Snippet: Nurr1 was revealed by Western blotting with a Nurr1 antibody (a gift from T. Perlmann) and an anti-rabbit antibody–horseradish peroxidase conjugate (Sigma).

    Techniques: Derivative Assay, Northern Blot, CTL Assay

    Effects of gene transfer on striatal dopaminergic innervation. The column on the left illustrates sections throughout the striatum of treated animals immunoreacted with an antibody raised against TH, to reveal the density of TH-immunopositive fibers in the striatum. The lesioned side is on the left, the control side, on the right. Notice that only animals injected with RAd-GDNF showed a protection of the striatal dopaminergic fibers. The graph shows the densitometric analysis of TH + fiber density in the striatum of rats treated with 1 × 10 8 IU of RAd-Shh, RAd-Gli-1, RAd-GDNF, or RAd-35. Degeneration of axonal terminals in the striatum after administration of 6-OHDA is not prevented following the injection of RAd-ShhN, RAd-Gli-1, or RAd-Nurr-1 (not shown). Only RAd-GDNF is able to protect TH + fibers significantly. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05. Scale bar, 1 mm.

    Journal: Molecular therapy : the journal of the American Society of Gene Therapy

    Article Title: Differentiation and Transcription Factor Gene Therapy in Experimental Parkinson's Disease: Sonic Hedgehog and Gli-1, but Not Nurr-1, Protect Nigrostriatal Cell Bodies from 6-OHDA-Induced Neurodegeneration

    doi: 10.1016/j.ymthe.2004.05.021

    Figure Lengend Snippet: Effects of gene transfer on striatal dopaminergic innervation. The column on the left illustrates sections throughout the striatum of treated animals immunoreacted with an antibody raised against TH, to reveal the density of TH-immunopositive fibers in the striatum. The lesioned side is on the left, the control side, on the right. Notice that only animals injected with RAd-GDNF showed a protection of the striatal dopaminergic fibers. The graph shows the densitometric analysis of TH + fiber density in the striatum of rats treated with 1 × 10 8 IU of RAd-Shh, RAd-Gli-1, RAd-GDNF, or RAd-35. Degeneration of axonal terminals in the striatum after administration of 6-OHDA is not prevented following the injection of RAd-ShhN, RAd-Gli-1, or RAd-Nurr-1 (not shown). Only RAd-GDNF is able to protect TH + fibers significantly. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05. Scale bar, 1 mm.

    Article Snippet: ShhN was detected using a mouse monoclonal antibody raised against the amino-terminal of Shh (University of Iowa Hybridoma Bank), Gli-1 and Nurr-1 were detected using rabbit polyclonal antibodies raised against the amino-terminal of Gli-1 or against Nurr-1 (Santa Cruz Biotechnology).

    Techniques: Injection

    Genomic structures of RAd-GDNF, RAd-ShhN, and RAd-Gli-1. Recombinant adenoviruses (RAd) were generated by homologous recombination after cotrans-fection into 293 cells of a shuttle expression plasmid encoding ShhN, Gli-1, or Nurr-1 together with the Ad5 genomic plasmid pJM17. The shuttle plasmid contained adenoviral DNA sequences encoding the left-end replication origin/packaging elements and the overlap – recombination region. Restriction patterns of ad-enoviral vectors digested with Hin dIII confirmed the presence of the transgenes. Identity of the transgenes was confirmed by Southern blot hybridization, using specific DIG-labeled probes. (a) The schematic structure of the new vectors described herein. (b, c) The characterization of RAd-GDNF, with (b) the restriction analysis and (c) the Southern blot hybridization indicating the presence of the expected transgene band (1.4 kb), in lanes 1 and 2. In b and c the lane numbers indicate 1, the shuttle vector used to construct the recombinant virus, pALGDNF (as positive control); 2, RAd-GDNF; 3, the Ad5 genomic plasmid pJM17 (as negative control); 4, molecular weight markers. (d, e) The characterization of RAd-ShhN, with (d) the restriction analysis and (e) the Southern blot hybridization indicating the presence of the band containing the transgene ShhN (0.9 kb), in lanes 1 and 2. In d and e the lane numbers indicate 1, RAd-ShhN; 2, the shuttle plasmid pALShhN (as positive control); 3, molecular weight markers. (f, g) The characterization of RAd-Gli-1, with (f) the restriction analysis and (g) the Southern blot hybridization indicating the presence of the band containing the transgene Gli-1, of approx 4.0 kb, in lanes 1 and 3. In f and g the lane numbers indicate MW, molecular weight markers; 1, RAd-Gli-1; 2, pJM17 (as negative control); 3, pALGli-1 (as positive control). The construction of Nurr-1 followed identical principles but has not been illustrated.

    Journal: Molecular therapy : the journal of the American Society of Gene Therapy

    Article Title: Differentiation and Transcription Factor Gene Therapy in Experimental Parkinson's Disease: Sonic Hedgehog and Gli-1, but Not Nurr-1, Protect Nigrostriatal Cell Bodies from 6-OHDA-Induced Neurodegeneration

    doi: 10.1016/j.ymthe.2004.05.021

    Figure Lengend Snippet: Genomic structures of RAd-GDNF, RAd-ShhN, and RAd-Gli-1. Recombinant adenoviruses (RAd) were generated by homologous recombination after cotrans-fection into 293 cells of a shuttle expression plasmid encoding ShhN, Gli-1, or Nurr-1 together with the Ad5 genomic plasmid pJM17. The shuttle plasmid contained adenoviral DNA sequences encoding the left-end replication origin/packaging elements and the overlap – recombination region. Restriction patterns of ad-enoviral vectors digested with Hin dIII confirmed the presence of the transgenes. Identity of the transgenes was confirmed by Southern blot hybridization, using specific DIG-labeled probes. (a) The schematic structure of the new vectors described herein. (b, c) The characterization of RAd-GDNF, with (b) the restriction analysis and (c) the Southern blot hybridization indicating the presence of the expected transgene band (1.4 kb), in lanes 1 and 2. In b and c the lane numbers indicate 1, the shuttle vector used to construct the recombinant virus, pALGDNF (as positive control); 2, RAd-GDNF; 3, the Ad5 genomic plasmid pJM17 (as negative control); 4, molecular weight markers. (d, e) The characterization of RAd-ShhN, with (d) the restriction analysis and (e) the Southern blot hybridization indicating the presence of the band containing the transgene ShhN (0.9 kb), in lanes 1 and 2. In d and e the lane numbers indicate 1, RAd-ShhN; 2, the shuttle plasmid pALShhN (as positive control); 3, molecular weight markers. (f, g) The characterization of RAd-Gli-1, with (f) the restriction analysis and (g) the Southern blot hybridization indicating the presence of the band containing the transgene Gli-1, of approx 4.0 kb, in lanes 1 and 3. In f and g the lane numbers indicate MW, molecular weight markers; 1, RAd-Gli-1; 2, pJM17 (as negative control); 3, pALGli-1 (as positive control). The construction of Nurr-1 followed identical principles but has not been illustrated.

    Article Snippet: ShhN was detected using a mouse monoclonal antibody raised against the amino-terminal of Shh (University of Iowa Hybridoma Bank), Gli-1 and Nurr-1 were detected using rabbit polyclonal antibodies raised against the amino-terminal of Gli-1 or against Nurr-1 (Santa Cruz Biotechnology).

    Techniques: Recombinant, Generated, Homologous Recombination, Expressing, Plasmid Preparation, Southern Blot, Hybridization, Labeling, Construct, Positive Control, Negative Control, Molecular Weight

    (a) In vitro bioactivity of RAd-ShhN, RAd-Gli-1, and RAd-Nurr-1. A schematic view of the mechanism of action of ShhN, and Gli-1, is shown. This illustrates the interaction of ShhN with Ptc and Smo, the release of the inhibition on Smo, and the eventual stimulation of activated Gli-1 translocation into the nucleus to activate further downstream target genes. (b) The bioactiv-ity of RAd-ShhN and RAd-Gli-1. HeLa cells were infected with RAd-ShhN, or RAd-Gli-1, or a negative control vector, at m.o.i. 200 IU/cell, and 48 h later cells were fixed and the proteins detected with specific primary antibodies and immunofluorescently labeled secondary antibodies. Most of the ShhN immunoreactivity highlighted the Golgi apparatus, compatible with the intracellular distribution of a secretory protein, while Gli-1 showed both a cytoplasmic and a nuclear localization, as expected from a transcription factor that has been shown to shuttle between the cytoplasm and the nucleus. Differentiation of the pluripotential cell line C3H10T1/2 into osteoblasts was induced upon infection with RAd-ShhN, or RAd-Gli-1, following infection at m.o.i. 200. Alkaline phosphatase (AP) activity was used as a marker for osteoblast differentiation. Both vectors induced AP activity, and the quantitative analysis of AP + cells is illustrated in (c). (d, e) The transcriptional activation mediated by RAd-Nurr-1 is shown. COS-7 cells were transfected with the reporter plasmid NBRE-Luciferase (d), containing the binding site for Nurr-1, and infected with RAd-Nurr-1 in sense orientation or RAd-Nurr-1 in antisense orientation (RAd-Nurr-1AS), used as negative control. (e) Luciferase activity was measured 48 h after. The transcription factor Nurr1 binds to the canonical NBRE domain and induces expression of luciferase. (f) COS-7 cells were infected with RAd-Nurr-1, or a negative control vector, at m.o.i. 200 IU/cell, and 48 h later cells were fixed and immunostained with antibodies recognizing Nurr1.

    Journal: Molecular therapy : the journal of the American Society of Gene Therapy

    Article Title: Differentiation and Transcription Factor Gene Therapy in Experimental Parkinson's Disease: Sonic Hedgehog and Gli-1, but Not Nurr-1, Protect Nigrostriatal Cell Bodies from 6-OHDA-Induced Neurodegeneration

    doi: 10.1016/j.ymthe.2004.05.021

    Figure Lengend Snippet: (a) In vitro bioactivity of RAd-ShhN, RAd-Gli-1, and RAd-Nurr-1. A schematic view of the mechanism of action of ShhN, and Gli-1, is shown. This illustrates the interaction of ShhN with Ptc and Smo, the release of the inhibition on Smo, and the eventual stimulation of activated Gli-1 translocation into the nucleus to activate further downstream target genes. (b) The bioactiv-ity of RAd-ShhN and RAd-Gli-1. HeLa cells were infected with RAd-ShhN, or RAd-Gli-1, or a negative control vector, at m.o.i. 200 IU/cell, and 48 h later cells were fixed and the proteins detected with specific primary antibodies and immunofluorescently labeled secondary antibodies. Most of the ShhN immunoreactivity highlighted the Golgi apparatus, compatible with the intracellular distribution of a secretory protein, while Gli-1 showed both a cytoplasmic and a nuclear localization, as expected from a transcription factor that has been shown to shuttle between the cytoplasm and the nucleus. Differentiation of the pluripotential cell line C3H10T1/2 into osteoblasts was induced upon infection with RAd-ShhN, or RAd-Gli-1, following infection at m.o.i. 200. Alkaline phosphatase (AP) activity was used as a marker for osteoblast differentiation. Both vectors induced AP activity, and the quantitative analysis of AP + cells is illustrated in (c). (d, e) The transcriptional activation mediated by RAd-Nurr-1 is shown. COS-7 cells were transfected with the reporter plasmid NBRE-Luciferase (d), containing the binding site for Nurr-1, and infected with RAd-Nurr-1 in sense orientation or RAd-Nurr-1 in antisense orientation (RAd-Nurr-1AS), used as negative control. (e) Luciferase activity was measured 48 h after. The transcription factor Nurr1 binds to the canonical NBRE domain and induces expression of luciferase. (f) COS-7 cells were infected with RAd-Nurr-1, or a negative control vector, at m.o.i. 200 IU/cell, and 48 h later cells were fixed and immunostained with antibodies recognizing Nurr1.

    Article Snippet: ShhN was detected using a mouse monoclonal antibody raised against the amino-terminal of Shh (University of Iowa Hybridoma Bank), Gli-1 and Nurr-1 were detected using rabbit polyclonal antibodies raised against the amino-terminal of Gli-1 or against Nurr-1 (Santa Cruz Biotechnology).

    Techniques: In Vitro, Inhibition, Translocation Assay, Infection, Negative Control, Plasmid Preparation, Labeling, Activity Assay, Marker, Activation Assay, Transfection, Luciferase, Binding Assay, Expressing

    Effects of gene transfer on substantia nigra dopaminergic neurons. Adenovirus-mediated gene transfer of 1 × 10 8 IU of (a, b) RAd-35, (c, d) RAd-GDNF, (e, f) RAd-ShhN, (g, h) RAd-Gli-1, or RAd-Nurr-1 (not illustrated) was tested against 6-OHDA-induced neurodegeneration of nigrostriatal cells retrogradely labeled with fluoro-gold. The side injected with 6-OHDA is shown on the left, and the control side is shown on the right. Injection of RAd-GDNF, RAd-ShhN, and RAd-Gli-1 protected a significant amount of nigrostriatal neurons compared to animals injected with the negative control vector RAd-35. Note the survival of large fluoro-gold + neurons in the ipsilateral site of animals injected with RAd-ShhN (e), RAd-Gli-1 (g), and RAd-GDNF (c) compared with RAd-35 (a). The quantitative analysis is shown in (i) and also indicates the analysis of the animals injected with RAd-Nurr-1. Survival of nigrostriatal neurons was expressed as a percentage of unlesioned contrala-teral neurons. (j) The area occupied by dopamine neurons’ cell bodies protected from degeneration after treatment with 1 × 10 8 IU of RAd-ShhN, RAd-Gli-1, or RAd-GDNF was quantified and expressed as a percentage of the neuron soma area in the contralateral site. RAd-GDNF, RAd-ShhN, and RAd-Gli-1 all protected cell body size compared with RAd-35. RAd-GDNF showed the strongest effect. Cell body protection by ShhN and Gli-1 was statistically significantly different from that of animals injected with RAd-35. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05; *** P ≤ 0.005.

    Journal: Molecular therapy : the journal of the American Society of Gene Therapy

    Article Title: Differentiation and Transcription Factor Gene Therapy in Experimental Parkinson's Disease: Sonic Hedgehog and Gli-1, but Not Nurr-1, Protect Nigrostriatal Cell Bodies from 6-OHDA-Induced Neurodegeneration

    doi: 10.1016/j.ymthe.2004.05.021

    Figure Lengend Snippet: Effects of gene transfer on substantia nigra dopaminergic neurons. Adenovirus-mediated gene transfer of 1 × 10 8 IU of (a, b) RAd-35, (c, d) RAd-GDNF, (e, f) RAd-ShhN, (g, h) RAd-Gli-1, or RAd-Nurr-1 (not illustrated) was tested against 6-OHDA-induced neurodegeneration of nigrostriatal cells retrogradely labeled with fluoro-gold. The side injected with 6-OHDA is shown on the left, and the control side is shown on the right. Injection of RAd-GDNF, RAd-ShhN, and RAd-Gli-1 protected a significant amount of nigrostriatal neurons compared to animals injected with the negative control vector RAd-35. Note the survival of large fluoro-gold + neurons in the ipsilateral site of animals injected with RAd-ShhN (e), RAd-Gli-1 (g), and RAd-GDNF (c) compared with RAd-35 (a). The quantitative analysis is shown in (i) and also indicates the analysis of the animals injected with RAd-Nurr-1. Survival of nigrostriatal neurons was expressed as a percentage of unlesioned contrala-teral neurons. (j) The area occupied by dopamine neurons’ cell bodies protected from degeneration after treatment with 1 × 10 8 IU of RAd-ShhN, RAd-Gli-1, or RAd-GDNF was quantified and expressed as a percentage of the neuron soma area in the contralateral site. RAd-GDNF, RAd-ShhN, and RAd-Gli-1 all protected cell body size compared with RAd-35. RAd-GDNF showed the strongest effect. Cell body protection by ShhN and Gli-1 was statistically significantly different from that of animals injected with RAd-35. The treatment groups were compared by repeated-measures ANOVA with post hoc Tukey or Dunnet multiple comparison test; * P ≤ 0.05; *** P ≤ 0.005.

    Article Snippet: ShhN was detected using a mouse monoclonal antibody raised against the amino-terminal of Shh (University of Iowa Hybridoma Bank), Gli-1 and Nurr-1 were detected using rabbit polyclonal antibodies raised against the amino-terminal of Gli-1 or against Nurr-1 (Santa Cruz Biotechnology).

    Techniques: Labeling, Injection, Negative Control, Plasmid Preparation

    Involvement of RXRα in the suppressive effect of PA024 on CYP11B2 mRNA expression and promoter activity. (A) and (B), effect of RXRα knockdown by its siRNA on the mRNA expression of CYP11B2 and NURR1. H295R cells transiently transfected with siRNA (negative control or RXRα) for 48 h were incubated either in the presence (10 μmol/L) or absence (control) of PA024 for 24 h and co-treated with 100 nmol/L Ang II for the last 6 h. Results are expressed as percentages of each Ang II. Data represent mean ± S.E.M. (n = 4). ** P

    Journal: PLoS ONE

    Article Title: Suppressive effects of RXR agonist PA024 on adrenal CYP11B2 expression, aldosterone secretion and blood pressure

    doi: 10.1371/journal.pone.0181055

    Figure Lengend Snippet: Involvement of RXRα in the suppressive effect of PA024 on CYP11B2 mRNA expression and promoter activity. (A) and (B), effect of RXRα knockdown by its siRNA on the mRNA expression of CYP11B2 and NURR1. H295R cells transiently transfected with siRNA (negative control or RXRα) for 48 h were incubated either in the presence (10 μmol/L) or absence (control) of PA024 for 24 h and co-treated with 100 nmol/L Ang II for the last 6 h. Results are expressed as percentages of each Ang II. Data represent mean ± S.E.M. (n = 4). ** P

    Article Snippet: Therefore, we first examined the effects of PA024 on the NURR1 and NGFIB mRNA expression levels in H295R cells.

    Techniques: Expressing, Activity Assay, Transfection, Negative Control, Incubation

    Possible involvement of NURR1 in the PA024-mediated suppression of CYP11B2 promoter activity via binding to the Ad5 element. (A) and (B), dose-response analyses of the NURR1 and NGFIB mRNA expression. H295R cells were treated with PA024 (indicated concentrations, 24h) and Ang II (100 nmol/L, 6 h). Data represent mean ± S.E.M. (n = 4), percent of control. *** P

    Journal: PLoS ONE

    Article Title: Suppressive effects of RXR agonist PA024 on adrenal CYP11B2 expression, aldosterone secretion and blood pressure

    doi: 10.1371/journal.pone.0181055

    Figure Lengend Snippet: Possible involvement of NURR1 in the PA024-mediated suppression of CYP11B2 promoter activity via binding to the Ad5 element. (A) and (B), dose-response analyses of the NURR1 and NGFIB mRNA expression. H295R cells were treated with PA024 (indicated concentrations, 24h) and Ang II (100 nmol/L, 6 h). Data represent mean ± S.E.M. (n = 4), percent of control. *** P

    Article Snippet: Therefore, we first examined the effects of PA024 on the NURR1 and NGFIB mRNA expression levels in H295R cells.

    Techniques: Activity Assay, Binding Assay, Expressing

    The impact of miR-145 overexpression and Nurr1 on the differentiation of NSCs induced by BMP2. NSCs were treated with 0 ng/ml of BMP2, or 20 ng/ml of BMP2, or 20 ng/ml of BMP2+NC, or 20 ng/ml of BMP2+miR-145 mimics, or 20 ng/ml of BMP2+miR-145 mimics+pcDNA-3.1,

    Journal: American Journal of Translational Research

    Article Title: BMP2 promotes the differentiation of neural stem cells into dopaminergic neurons in vitro via miR-145-mediated upregulation of Nurr1 expression

    doi:

    Figure Lengend Snippet: The impact of miR-145 overexpression and Nurr1 on the differentiation of NSCs induced by BMP2. NSCs were treated with 0 ng/ml of BMP2, or 20 ng/ml of BMP2, or 20 ng/ml of BMP2+NC, or 20 ng/ml of BMP2+miR-145 mimics, or 20 ng/ml of BMP2+miR-145 mimics+pcDNA-3.1,

    Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membrane was incubated with primary antibody Nurr1 (1:1000, Abcam) and β-atcin (1:1000, Abcam, Cambridge, UK) for 3 h at room temperature.

    Techniques: Over Expression

    The effect of miR-145 inhibitor and si-Nurr1 on the differentiation of NSCs into DA neurons. NSCs were transfected with NC, or miR-145 inhibitor, or miR-145 inhibitor and si-NC, or miR-145 inhibitor and si-Nurr1. Then differentiation liquid without BMP2

    Journal: American Journal of Translational Research

    Article Title: BMP2 promotes the differentiation of neural stem cells into dopaminergic neurons in vitro via miR-145-mediated upregulation of Nurr1 expression

    doi:

    Figure Lengend Snippet: The effect of miR-145 inhibitor and si-Nurr1 on the differentiation of NSCs into DA neurons. NSCs were transfected with NC, or miR-145 inhibitor, or miR-145 inhibitor and si-NC, or miR-145 inhibitor and si-Nurr1. Then differentiation liquid without BMP2

    Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membrane was incubated with primary antibody Nurr1 (1:1000, Abcam) and β-atcin (1:1000, Abcam, Cambridge, UK) for 3 h at room temperature.

    Techniques: Transfection

    miR-145 impaired the expression of Nurr1 through targeting the 3’-UTR sequences of Nurr1. A. The putative miR-145 binding site within the human Nurr1 3’-UTR (WT) and the 3’-UTR mutated sequence (MU) were showed; B. Luciferase reporter

    Journal: American Journal of Translational Research

    Article Title: BMP2 promotes the differentiation of neural stem cells into dopaminergic neurons in vitro via miR-145-mediated upregulation of Nurr1 expression

    doi:

    Figure Lengend Snippet: miR-145 impaired the expression of Nurr1 through targeting the 3’-UTR sequences of Nurr1. A. The putative miR-145 binding site within the human Nurr1 3’-UTR (WT) and the 3’-UTR mutated sequence (MU) were showed; B. Luciferase reporter

    Article Snippet: After blocked with 5% non-fat milk for 2 h at room temperature, the membrane was incubated with primary antibody Nurr1 (1:1000, Abcam) and β-atcin (1:1000, Abcam, Cambridge, UK) for 3 h at room temperature.

    Techniques: Expressing, Binding Assay, Sequencing, Luciferase

    miR-381-3p inhibition provides protective effects by targeting nurr1. a Immunofluorescence staining for the Ki-67 antibody in IEC-6 cells for proliferation analysis. IEC-6 cells were infected with si-Nurr1 or si-control, transfected with the ant-381 or the ant-NC, and then incubated in H/R conditions for 6/6 h, respectively. Scale bar = 100 μm, n = 6. b and c Caco-2 cells were co-transfected with ant-381, si-Nurr1 or the corresponding negative control, and then exposed to H/R conditions for 12/6 h, respectively. b Effect of the ant-381 on TEER, n = 3. c Effect of the ant-381 on FITC-dextran intestinal epithelial permeability, n = 3. d-h IEC-6 cells were co-transfected with ant-381, si-Nurr1 or the corresponding negative control, and then incubated in H/R for 6/6 h, respectively. Representative western blot showing nurr1, p21, occludin, ZO-1 protein expression, n = 3. * P

    Journal: Cell Death & Disease

    Article Title: miR-381-3p knockdown improves intestinal epithelial proliferation and barrier function after intestinal ischemia/reperfusion injury by targeting nurr1

    doi: 10.1038/s41419-018-0450-z

    Figure Lengend Snippet: miR-381-3p inhibition provides protective effects by targeting nurr1. a Immunofluorescence staining for the Ki-67 antibody in IEC-6 cells for proliferation analysis. IEC-6 cells were infected with si-Nurr1 or si-control, transfected with the ant-381 or the ant-NC, and then incubated in H/R conditions for 6/6 h, respectively. Scale bar = 100 μm, n = 6. b and c Caco-2 cells were co-transfected with ant-381, si-Nurr1 or the corresponding negative control, and then exposed to H/R conditions for 12/6 h, respectively. b Effect of the ant-381 on TEER, n = 3. c Effect of the ant-381 on FITC-dextran intestinal epithelial permeability, n = 3. d-h IEC-6 cells were co-transfected with ant-381, si-Nurr1 or the corresponding negative control, and then incubated in H/R for 6/6 h, respectively. Representative western blot showing nurr1, p21, occludin, ZO-1 protein expression, n = 3. * P

    Article Snippet: Western blotting was performed with primary antibodies against nurr1, p21 (Abcam, Cambridge, UK), ZO-1, occludin (Proteintech, Wuhan, China), and β-actin (ZSGB-BIO, Beijing, China) and horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB-BIO, Beijing, China).

    Techniques: Inhibition, Immunofluorescence, Staining, Infection, Transfection, Incubation, Negative Control, Permeability, Western Blot, Expressing

    miR-381-3p, nurr1 and tight junction protein expression levels in the ischemic intestine of clinical patients. a qRT-PCR showing miR-381-3p expression in normal and ischemic human intestines, n = 6. b The association between miR-381-3p expression and Nurr1 protein expression ( r 2 = 0.8052, p = 0.0153). c and d Representative western blot showing nurr1, p21, occludin or ZO-1 protein expression in the intestine, n = 3. * P

    Journal: Cell Death & Disease

    Article Title: miR-381-3p knockdown improves intestinal epithelial proliferation and barrier function after intestinal ischemia/reperfusion injury by targeting nurr1

    doi: 10.1038/s41419-018-0450-z

    Figure Lengend Snippet: miR-381-3p, nurr1 and tight junction protein expression levels in the ischemic intestine of clinical patients. a qRT-PCR showing miR-381-3p expression in normal and ischemic human intestines, n = 6. b The association between miR-381-3p expression and Nurr1 protein expression ( r 2 = 0.8052, p = 0.0153). c and d Representative western blot showing nurr1, p21, occludin or ZO-1 protein expression in the intestine, n = 3. * P

    Article Snippet: Western blotting was performed with primary antibodies against nurr1, p21 (Abcam, Cambridge, UK), ZO-1, occludin (Proteintech, Wuhan, China), and β-actin (ZSGB-BIO, Beijing, China) and horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB-BIO, Beijing, China).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    Nurr1 regulates intestinal restoration after I/R injury. a Mice were subjected to 45 min of intestinal ischemia followed by 0–16 h of reperfusion or sham surgery. I, ischemia; R, reperfusion; representative western blot showing nurr1 protein expression in the intestinal tissue lysates ( n = 3 per group, ** P

    Journal: Cell Death & Disease

    Article Title: miR-381-3p knockdown improves intestinal epithelial proliferation and barrier function after intestinal ischemia/reperfusion injury by targeting nurr1

    doi: 10.1038/s41419-018-0450-z

    Figure Lengend Snippet: Nurr1 regulates intestinal restoration after I/R injury. a Mice were subjected to 45 min of intestinal ischemia followed by 0–16 h of reperfusion or sham surgery. I, ischemia; R, reperfusion; representative western blot showing nurr1 protein expression in the intestinal tissue lysates ( n = 3 per group, ** P

    Article Snippet: Western blotting was performed with primary antibodies against nurr1, p21 (Abcam, Cambridge, UK), ZO-1, occludin (Proteintech, Wuhan, China), and β-actin (ZSGB-BIO, Beijing, China) and horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB-BIO, Beijing, China).

    Techniques: Mouse Assay, Western Blot, Expressing

    miR-381-3p negatively regulates nurr1 expression. a The miR-381-3p target sequence in the nurr1 3ʹ-UTR is conserved across various species. b The WT nurr1 3ʹ-UTR and the MT nurr1 3ʹ-UTR in the luciferase constructs. BS, binding site. c Caco-2 cells were infected with miR-381 or miR-NC and WT nurr1 3ʹ-UTR or MT nurr1 3ʹ-UTR, n = 3. d-g Caco-2 or IEC-6 cells were transfected with the ant-381 or the ant-NC. d Representative western blot showing nurr1 protein expression in Caco-2 cells, n = 3. e qRT-PCR showing nurr1 mRNA expression in Caco-2 cells, n = 6. f Representative western blot showing nurr1 protein expression in IEC-6 cells, n = 3. g qRT-PCR showing nurr1 mRNA expression in IEC-6 cells, n = 6. * P

    Journal: Cell Death & Disease

    Article Title: miR-381-3p knockdown improves intestinal epithelial proliferation and barrier function after intestinal ischemia/reperfusion injury by targeting nurr1

    doi: 10.1038/s41419-018-0450-z

    Figure Lengend Snippet: miR-381-3p negatively regulates nurr1 expression. a The miR-381-3p target sequence in the nurr1 3ʹ-UTR is conserved across various species. b The WT nurr1 3ʹ-UTR and the MT nurr1 3ʹ-UTR in the luciferase constructs. BS, binding site. c Caco-2 cells were infected with miR-381 or miR-NC and WT nurr1 3ʹ-UTR or MT nurr1 3ʹ-UTR, n = 3. d-g Caco-2 or IEC-6 cells were transfected with the ant-381 or the ant-NC. d Representative western blot showing nurr1 protein expression in Caco-2 cells, n = 3. e qRT-PCR showing nurr1 mRNA expression in Caco-2 cells, n = 6. f Representative western blot showing nurr1 protein expression in IEC-6 cells, n = 3. g qRT-PCR showing nurr1 mRNA expression in IEC-6 cells, n = 6. * P

    Article Snippet: Western blotting was performed with primary antibodies against nurr1, p21 (Abcam, Cambridge, UK), ZO-1, occludin (Proteintech, Wuhan, China), and β-actin (ZSGB-BIO, Beijing, China) and horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB-BIO, Beijing, China).

    Techniques: Expressing, Sequencing, Luciferase, Construct, Binding Assay, Infection, Transfection, Western Blot, Quantitative RT-PCR

    miR-381-3p inhibition improves intestinal barrier restoration after I/R injury in mice. The mice were divided into the following four groups: a sham + LNA-NC group, a sham + LNA-381 group, an I/R + LNA-NC group, an I/R + LNA-381 group ( n = 8 per group). LNA-381 or LNA-NC (2 mg/kg) was administered by caudal vein injection at 12 h before surgery. The I/R times were 45/240 min, respectively. a qRT-PCR showing miR-381-3p expression, n = 8. b and c Immunohistochemical staining for the Ki-67 antibody in intestinal tissues for proliferation analysis. Scale bar = 50 μm, n = 6. d Serum d -lactate levels, n = 8. e FITC-dextran intestinal epithelial paracellular permeability, n = 8. f and g Representative images of H E-stained intestinal sections of mice from the above four groups. Intestinal injury was scored histopathologically (Chiu’s score) according to a scoring system. Scale bar = 100 μm, n = 8. h-l Representative western blot showing nurr1, p21, occludin or ZO-1 protein expression in intestinal tissue, n = 3. m The survival rate of the mice, n = 15. * P

    Journal: Cell Death & Disease

    Article Title: miR-381-3p knockdown improves intestinal epithelial proliferation and barrier function after intestinal ischemia/reperfusion injury by targeting nurr1

    doi: 10.1038/s41419-018-0450-z

    Figure Lengend Snippet: miR-381-3p inhibition improves intestinal barrier restoration after I/R injury in mice. The mice were divided into the following four groups: a sham + LNA-NC group, a sham + LNA-381 group, an I/R + LNA-NC group, an I/R + LNA-381 group ( n = 8 per group). LNA-381 or LNA-NC (2 mg/kg) was administered by caudal vein injection at 12 h before surgery. The I/R times were 45/240 min, respectively. a qRT-PCR showing miR-381-3p expression, n = 8. b and c Immunohistochemical staining for the Ki-67 antibody in intestinal tissues for proliferation analysis. Scale bar = 50 μm, n = 6. d Serum d -lactate levels, n = 8. e FITC-dextran intestinal epithelial paracellular permeability, n = 8. f and g Representative images of H E-stained intestinal sections of mice from the above four groups. Intestinal injury was scored histopathologically (Chiu’s score) according to a scoring system. Scale bar = 100 μm, n = 8. h-l Representative western blot showing nurr1, p21, occludin or ZO-1 protein expression in intestinal tissue, n = 3. m The survival rate of the mice, n = 15. * P

    Article Snippet: Western blotting was performed with primary antibodies against nurr1, p21 (Abcam, Cambridge, UK), ZO-1, occludin (Proteintech, Wuhan, China), and β-actin (ZSGB-BIO, Beijing, China) and horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB-BIO, Beijing, China).

    Techniques: Inhibition, Mouse Assay, Injection, Quantitative RT-PCR, Expressing, Immunohistochemistry, Staining, Permeability, Western Blot

    miR-381-3p inhibition promotes intestinal epithelial restoration after H/R injury. a Immunofluorescence staining for the Ki-67 antibody in IEC-6 cells for proliferation analysis. IEC-6 cells were infected with the ant-381 or the ant-NC for 36 h and then incubated under H/R conditions for 6/6 h, respectively. Scale bar = 100 μm, n = 6. b and c Caco-2 cells were transfected with the ant-381 or the ant-NC for 36 h and then exposed to hypoxic conditions for 12 h before being exposed to normoxic conditions for 6 h. b Effect of the ant-381 on TEER, n = 3. c Changes in FITC-dextran intestinal epithelial permeability in response to the ant-381, n = 3. d-h IEC-6 cells were infected with the ant-381 or the ant-NC for 36 h and then incubated under H/R conditions for 6/6 h, respectively. Representative western blot showing nurr1, p21, occludin, ZO-1 protein expression, n = 3. * P

    Journal: Cell Death & Disease

    Article Title: miR-381-3p knockdown improves intestinal epithelial proliferation and barrier function after intestinal ischemia/reperfusion injury by targeting nurr1

    doi: 10.1038/s41419-018-0450-z

    Figure Lengend Snippet: miR-381-3p inhibition promotes intestinal epithelial restoration after H/R injury. a Immunofluorescence staining for the Ki-67 antibody in IEC-6 cells for proliferation analysis. IEC-6 cells were infected with the ant-381 or the ant-NC for 36 h and then incubated under H/R conditions for 6/6 h, respectively. Scale bar = 100 μm, n = 6. b and c Caco-2 cells were transfected with the ant-381 or the ant-NC for 36 h and then exposed to hypoxic conditions for 12 h before being exposed to normoxic conditions for 6 h. b Effect of the ant-381 on TEER, n = 3. c Changes in FITC-dextran intestinal epithelial permeability in response to the ant-381, n = 3. d-h IEC-6 cells were infected with the ant-381 or the ant-NC for 36 h and then incubated under H/R conditions for 6/6 h, respectively. Representative western blot showing nurr1, p21, occludin, ZO-1 protein expression, n = 3. * P

    Article Snippet: Western blotting was performed with primary antibodies against nurr1, p21 (Abcam, Cambridge, UK), ZO-1, occludin (Proteintech, Wuhan, China), and β-actin (ZSGB-BIO, Beijing, China) and horseradish peroxidase (HRP)-conjugated secondary antibodies (ZSGB-BIO, Beijing, China).

    Techniques: Inhibition, Immunofluorescence, Staining, Infection, Incubation, Transfection, Permeability, Western Blot, Expressing

    CREB regulates Nurr1 induction by NMDA. Neurons cultured in K5 were treated (or non-treated; NT ) with NMDA (100 μ m ) or K25 at 2 DIV. A , phosphorylation of CREB ( pCREB ) and total CREB were determined by Western blot at the indicated times. Data

    Journal: The Journal of Biological Chemistry

    Article Title: Nurr1 Protein Is Required for N-Methyl-d-aspartic Acid (NMDA) Receptor-mediated Neuronal Survival

    doi: 10.1074/jbc.M111.272427

    Figure Lengend Snippet: CREB regulates Nurr1 induction by NMDA. Neurons cultured in K5 were treated (or non-treated; NT ) with NMDA (100 μ m ) or K25 at 2 DIV. A , phosphorylation of CREB ( pCREB ) and total CREB were determined by Western blot at the indicated times. Data

    Article Snippet: 2.5 μg of soluble chromatin were co-immunoprecipitated with 0.5 μl of phospho-Ser133 -CREB antibody (Cell Signaling), Nurr1 antibody (Abcam), or rabbit IgG (Santa Cruz Biotechnology).

    Techniques: Cell Culture, Western Blot

    Nurr1 regulates NMDA-mediated increase in BDNF levels and has similar expression pattern to neurotrophin during postnatal cerebellum development. A , CGCs were plated in the presence or absence of the indicated shRNA and treated (or non-treated; NT ) at

    Journal: The Journal of Biological Chemistry

    Article Title: Nurr1 Protein Is Required for N-Methyl-d-aspartic Acid (NMDA) Receptor-mediated Neuronal Survival

    doi: 10.1074/jbc.M111.272427

    Figure Lengend Snippet: Nurr1 regulates NMDA-mediated increase in BDNF levels and has similar expression pattern to neurotrophin during postnatal cerebellum development. A , CGCs were plated in the presence or absence of the indicated shRNA and treated (or non-treated; NT ) at

    Article Snippet: 2.5 μg of soluble chromatin were co-immunoprecipitated with 0.5 μl of phospho-Ser133 -CREB antibody (Cell Signaling), Nurr1 antibody (Abcam), or rabbit IgG (Santa Cruz Biotechnology).

    Techniques: Expressing, shRNA

    Nr4a Family members are up-regulated in response to NMDA, but only Nurr1 protein levels are increased. CGCs were plated in 25 m m KCl ( NT ) or K5. At 2 DIV, K5 cells were treated or not with NMDA (100 μ m ) ( N ) or K25 ( K ). A and B , at the indicated

    Journal: The Journal of Biological Chemistry

    Article Title: Nurr1 Protein Is Required for N-Methyl-d-aspartic Acid (NMDA) Receptor-mediated Neuronal Survival

    doi: 10.1074/jbc.M111.272427

    Figure Lengend Snippet: Nr4a Family members are up-regulated in response to NMDA, but only Nurr1 protein levels are increased. CGCs were plated in 25 m m KCl ( NT ) or K5. At 2 DIV, K5 cells were treated or not with NMDA (100 μ m ) ( N ) or K25 ( K ). A and B , at the indicated

    Article Snippet: 2.5 μg of soluble chromatin were co-immunoprecipitated with 0.5 μl of phospho-Ser133 -CREB antibody (Cell Signaling), Nurr1 antibody (Abcam), or rabbit IgG (Santa Cruz Biotechnology).

    Techniques:

    Nurr1 expression is mainly restricted to nodulus lobe. Sagittal sections of cerebellum from P9, P14, and P21 rats were obtained and subjected to Nurr1 immunohistochemistry. Nurr1-increased expression during the different postnatal days was clearly evident

    Journal: The Journal of Biological Chemistry

    Article Title: Nurr1 Protein Is Required for N-Methyl-d-aspartic Acid (NMDA) Receptor-mediated Neuronal Survival

    doi: 10.1074/jbc.M111.272427

    Figure Lengend Snippet: Nurr1 expression is mainly restricted to nodulus lobe. Sagittal sections of cerebellum from P9, P14, and P21 rats were obtained and subjected to Nurr1 immunohistochemistry. Nurr1-increased expression during the different postnatal days was clearly evident

    Article Snippet: 2.5 μg of soluble chromatin were co-immunoprecipitated with 0.5 μl of phospho-Ser133 -CREB antibody (Cell Signaling), Nurr1 antibody (Abcam), or rabbit IgG (Santa Cruz Biotechnology).

    Techniques: Expressing, Immunohistochemistry

    Nurr1 participates in NMDA neuroprotective effect. CGCs were plated in the presence of shRNA against Nurr1 (Nurr1_1) or a scrambled shRNA (scNurr1) as described under ”Experimental Procedures.“ At 2 DIV, cells were treated (or non-treated;

    Journal: The Journal of Biological Chemistry

    Article Title: Nurr1 Protein Is Required for N-Methyl-d-aspartic Acid (NMDA) Receptor-mediated Neuronal Survival

    doi: 10.1074/jbc.M111.272427

    Figure Lengend Snippet: Nurr1 participates in NMDA neuroprotective effect. CGCs were plated in the presence of shRNA against Nurr1 (Nurr1_1) or a scrambled shRNA (scNurr1) as described under ”Experimental Procedures.“ At 2 DIV, cells were treated (or non-treated;

    Article Snippet: 2.5 μg of soluble chromatin were co-immunoprecipitated with 0.5 μl of phospho-Ser133 -CREB antibody (Cell Signaling), Nurr1 antibody (Abcam), or rabbit IgG (Santa Cruz Biotechnology).

    Techniques: shRNA

    Functional effects of Nurr1 overexpression on AD‐related pathology in 5XFAD mice. (a) Injection of lentiviral vectors expressing Nurr1 significantly increases expression of Nurr1 in the subiculum of both WT and 5XFAD mice ( n = 3). *** p

    Journal: Aging Cell

    Article Title: Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model, et al. Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model

    doi: 10.1111/acel.12866

    Figure Lengend Snippet: Functional effects of Nurr1 overexpression on AD‐related pathology in 5XFAD mice. (a) Injection of lentiviral vectors expressing Nurr1 significantly increases expression of Nurr1 in the subiculum of both WT and 5XFAD mice ( n = 3). *** p

    Article Snippet: After blocking, membranes were incubated with the following primary antibodies: rabbit anti‐Nurr1 (1:1,000) and rabbit anti‐β‐actin (Abcam, Cambridge, MA; 1:5,000).

    Techniques: Functional Assay, Over Expression, Mouse Assay, Injection, Expressing

    Nurr1 expression in Aβ accumulating cells in 5XFAD mouse brains and in the postmortem brains of human AD patients. Nurr1 and 4G8 double labeling in the subiculum (a) and in the cerebral cortex (b) of 2‐, 4‐ and 6‐month‐old 5XFAD mice ( n = 4–5). (c) Immunostaining with antibodies against Nurr1 and EAAC1 in the frontal cortex and in the subiculum of 5XFAD mice at 2 months of age. (d) Nurr1 and 4G8 double labeling of the hippocampus of postmortem human AD brains. Analysis of Nurr1‐positive cells in the hippocampus (e) and frontal cortex (f) of control and AD human brains. All brain tissues were counterstained with DAPI. (g) Western blot analysis and quantification of Nurr1 expression in the hippocampal formation, superior frontal cortex, and substantia nigra of normal and AD patient brains. (h) Classification of Nurr1‐expressing cells according to Aβ expression in the subiculum of 5XFAD mice. * p

    Journal: Aging Cell

    Article Title: Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model, et al. Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model

    doi: 10.1111/acel.12866

    Figure Lengend Snippet: Nurr1 expression in Aβ accumulating cells in 5XFAD mouse brains and in the postmortem brains of human AD patients. Nurr1 and 4G8 double labeling in the subiculum (a) and in the cerebral cortex (b) of 2‐, 4‐ and 6‐month‐old 5XFAD mice ( n = 4–5). (c) Immunostaining with antibodies against Nurr1 and EAAC1 in the frontal cortex and in the subiculum of 5XFAD mice at 2 months of age. (d) Nurr1 and 4G8 double labeling of the hippocampus of postmortem human AD brains. Analysis of Nurr1‐positive cells in the hippocampus (e) and frontal cortex (f) of control and AD human brains. All brain tissues were counterstained with DAPI. (g) Western blot analysis and quantification of Nurr1 expression in the hippocampal formation, superior frontal cortex, and substantia nigra of normal and AD patient brains. (h) Classification of Nurr1‐expressing cells according to Aβ expression in the subiculum of 5XFAD mice. * p

    Article Snippet: After blocking, membranes were incubated with the following primary antibodies: rabbit anti‐Nurr1 (1:1,000) and rabbit anti‐β‐actin (Abcam, Cambridge, MA; 1:5,000).

    Techniques: Expressing, Labeling, Mouse Assay, Immunostaining, Western Blot

    Treatment with AQ inhibits Aβ‐mediated pathology and improves cognitive function in 5XFAD mice. (a) Nurr1 agonist significantly reduced 4G8‐positive plaques in the subiculum of 5XFAD mice ( n = 10). Scale bar = 50 μm. (b) Representative immunoblot image and quantification of IDE expression levels from AQ‐treated SH‐SY5Y cells. * p

    Journal: Aging Cell

    Article Title: Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model, et al. Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model

    doi: 10.1111/acel.12866

    Figure Lengend Snippet: Treatment with AQ inhibits Aβ‐mediated pathology and improves cognitive function in 5XFAD mice. (a) Nurr1 agonist significantly reduced 4G8‐positive plaques in the subiculum of 5XFAD mice ( n = 10). Scale bar = 50 μm. (b) Representative immunoblot image and quantification of IDE expression levels from AQ‐treated SH‐SY5Y cells. * p

    Article Snippet: After blocking, membranes were incubated with the following primary antibodies: rabbit anti‐Nurr1 (1:1,000) and rabbit anti‐β‐actin (Abcam, Cambridge, MA; 1:5,000).

    Techniques: Mouse Assay, Expressing

    Nurr1 is cell type‐specific expressed in glutamatergic neurons in the brains of wild‐type littermates and 5XFAD mice. (a, b) Double labeling of WT and 5XFAD mice brains (4 months of age ( n = 4–5)) with NeuN‐ and Nurr1‐specific antibodies. White circles in B indicate NeuN‐negative and Nurr1‐positive glial cells. Scale bar = 20 μm. (c) Increase in astrogliosis in the subiculum of 5XFAD as shown by staining for GFAP, an astrocyte marker. Double labeling with Nurr1 and with astrocyte marker GFAP antibodies (d) or with microglia marker Iba‐1 antibody (e) in the subiculum of 5XFAD mice. Immunostaining with antibodies against Nurr1 and EAAC1 (or vGlut1), a marker for glutamatergic neurons, in the hippocampus: e, CA1; f, CA3; g, subiculum; h, dentate gyrus; b, subiculum; c, CA1; d, CA3; e, dentate gyrus (DG). DAPI was used to stain the nuclei. Scale bar = 50 μm (a–e and g), Scale bar = 100 μm (f and h)

    Journal: Aging Cell

    Article Title: Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model, et al. Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model

    doi: 10.1111/acel.12866

    Figure Lengend Snippet: Nurr1 is cell type‐specific expressed in glutamatergic neurons in the brains of wild‐type littermates and 5XFAD mice. (a, b) Double labeling of WT and 5XFAD mice brains (4 months of age ( n = 4–5)) with NeuN‐ and Nurr1‐specific antibodies. White circles in B indicate NeuN‐negative and Nurr1‐positive glial cells. Scale bar = 20 μm. (c) Increase in astrogliosis in the subiculum of 5XFAD as shown by staining for GFAP, an astrocyte marker. Double labeling with Nurr1 and with astrocyte marker GFAP antibodies (d) or with microglia marker Iba‐1 antibody (e) in the subiculum of 5XFAD mice. Immunostaining with antibodies against Nurr1 and EAAC1 (or vGlut1), a marker for glutamatergic neurons, in the hippocampus: e, CA1; f, CA3; g, subiculum; h, dentate gyrus; b, subiculum; c, CA1; d, CA3; e, dentate gyrus (DG). DAPI was used to stain the nuclei. Scale bar = 50 μm (a–e and g), Scale bar = 100 μm (f and h)

    Article Snippet: After blocking, membranes were incubated with the following primary antibodies: rabbit anti‐Nurr1 (1:1,000) and rabbit anti‐β‐actin (Abcam, Cambridge, MA; 1:5,000).

    Techniques: Mouse Assay, Labeling, Staining, Marker, Immunostaining

    Functional effects of Nurr1 knockdown on histopathological manifestations of AD in 5XFAD mice. (a) Injection of sh‐Nurr1 lentivirus significantly reduces expression of Nurr1 in the subiculum of both WT and 5XFAD mice ( n = 3). *** p

    Journal: Aging Cell

    Article Title: Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model, et al. Nurr1 (NR4A2) regulates Alzheimer’s disease‐related pathogenesis and cognitive function in the 5XFAD mouse model

    doi: 10.1111/acel.12866

    Figure Lengend Snippet: Functional effects of Nurr1 knockdown on histopathological manifestations of AD in 5XFAD mice. (a) Injection of sh‐Nurr1 lentivirus significantly reduces expression of Nurr1 in the subiculum of both WT and 5XFAD mice ( n = 3). *** p

    Article Snippet: After blocking, membranes were incubated with the following primary antibodies: rabbit anti‐Nurr1 (1:1,000) and rabbit anti‐β‐actin (Abcam, Cambridge, MA; 1:5,000).

    Techniques: Functional Assay, Mouse Assay, Injection, Expressing

    ( A ) Representative images showing the co-localization of Nurr1 and TH positive cells expression in the VTA. Bars = 25 μm. ( B – D ) Quantitative analysis of TH + , Nurr1 + and Nurr1 + -TH + positive cells. The data are shown as mean ± SEM, * P

    Journal: International Journal of Molecular Sciences

    Article Title: Dysregulation of Dopaminergic Regulatory Factors TH, Nurr1, and Pitx3 in the Ventral Tegmental Area Associated with Neuronal Injury Induced by Chronic Morphine Dependence

    doi: 10.3390/ijms20020250

    Figure Lengend Snippet: ( A ) Representative images showing the co-localization of Nurr1 and TH positive cells expression in the VTA. Bars = 25 μm. ( B – D ) Quantitative analysis of TH + , Nurr1 + and Nurr1 + -TH + positive cells. The data are shown as mean ± SEM, * P

    Article Snippet: Reagents Rabbit monoclonal anti-TH antibody, mouse monoclonal anti-Nurr1 antibody, mouse monoclonal anti-Pitx3 antibody and anti-β-actin antibody were purchased from Abcam (Cambridge, UK).

    Techniques: Expressing

    Western blot analysis of Nurr1 expression in the VTA. The relative level of Nurr1 in the VTA after one week of morphine exposure increased compared with the control group ( P

    Journal: International Journal of Molecular Sciences

    Article Title: Dysregulation of Dopaminergic Regulatory Factors TH, Nurr1, and Pitx3 in the Ventral Tegmental Area Associated with Neuronal Injury Induced by Chronic Morphine Dependence

    doi: 10.3390/ijms20020250

    Figure Lengend Snippet: Western blot analysis of Nurr1 expression in the VTA. The relative level of Nurr1 in the VTA after one week of morphine exposure increased compared with the control group ( P

    Article Snippet: Reagents Rabbit monoclonal anti-TH antibody, mouse monoclonal anti-Nurr1 antibody, mouse monoclonal anti-Pitx3 antibody and anti-β-actin antibody were purchased from Abcam (Cambridge, UK).

    Techniques: Western Blot, Expressing

    Exogenous expression of A53T α-syn suppresses the nuclear accumulation of Nurr1 and the expression of Nurr1-controlled dopaminergic genes ( A ) Nurr1 (green) and TH (red) co-staining in the SNC coronal sections of 1-month-old of nTg , A53T , and DOX-treated A53T ( A53T/DOX ) mice. Asterisks indicate TH-positive neurons with severe loss of Nurr1 staining. Arrows mark the mDA neurons with intensified Nurr1 staining. Arrowheads point to non-DA cells stained with Nurr1. Scale bar: 20 μm ( B ) The intensity of Nurr1 signals in the nucleus of SNC mDA neurons from 1-month-old nTg , A53T , and A53T/DOX mice (n ≥ 3 animals per genotype; ≥ 40 neurons per animal). Data were presented as Mean ± SEM. ***p

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Conditional Expression of Parkinson disease-related Mutant α-synuclein in the Midbrain Dopaminergic Neurons causes Progressive Neurodegeneration and Degradation of Transcription Factor Nuclear Receptor Related 1

    doi: 10.1523/JNEUROSCI.1731-12.2012

    Figure Lengend Snippet: Exogenous expression of A53T α-syn suppresses the nuclear accumulation of Nurr1 and the expression of Nurr1-controlled dopaminergic genes ( A ) Nurr1 (green) and TH (red) co-staining in the SNC coronal sections of 1-month-old of nTg , A53T , and DOX-treated A53T ( A53T/DOX ) mice. Asterisks indicate TH-positive neurons with severe loss of Nurr1 staining. Arrows mark the mDA neurons with intensified Nurr1 staining. Arrowheads point to non-DA cells stained with Nurr1. Scale bar: 20 μm ( B ) The intensity of Nurr1 signals in the nucleus of SNC mDA neurons from 1-month-old nTg , A53T , and A53T/DOX mice (n ≥ 3 animals per genotype; ≥ 40 neurons per animal). Data were presented as Mean ± SEM. ***p

    Article Snippet: Cells were then double-labeled with primary antibodies against TH (Santa Cruz; mouse monoclonal; 1:1000) and Nurr1 (Sigma; rabbit polyclonal; 1:1000) overnight at 4°C in a humidified chamber.

    Techniques: Expressing, Staining, Mouse Assay, Multiple Displacement Amplification

    A53T α-syn promotes proteasome-dependent degradation of Nurr1 and inhibition of proteasome activities ameliorates A53T α-syn-induced loss of mDA neurons ( A ) Expression of endogenous Nurr1 protein in the HEK293 cells transfected with human WT or A53T α-syn cDNA. The transfected HEK293 cells were treated with either DMSO or 10 μM MG132 for 12hr before being lysed for Western blot analysis. β-actin was used as loading control. ( B ) Bar graph summarizes the expression of endogenous Nurr1 in α-syn-transfected HEK293 cells treated with DMSO or MG132, from three independent experiments. Data were presented as Mean ± SEM. *p

    Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

    Article Title: Conditional Expression of Parkinson disease-related Mutant α-synuclein in the Midbrain Dopaminergic Neurons causes Progressive Neurodegeneration and Degradation of Transcription Factor Nuclear Receptor Related 1

    doi: 10.1523/JNEUROSCI.1731-12.2012

    Figure Lengend Snippet: A53T α-syn promotes proteasome-dependent degradation of Nurr1 and inhibition of proteasome activities ameliorates A53T α-syn-induced loss of mDA neurons ( A ) Expression of endogenous Nurr1 protein in the HEK293 cells transfected with human WT or A53T α-syn cDNA. The transfected HEK293 cells were treated with either DMSO or 10 μM MG132 for 12hr before being lysed for Western blot analysis. β-actin was used as loading control. ( B ) Bar graph summarizes the expression of endogenous Nurr1 in α-syn-transfected HEK293 cells treated with DMSO or MG132, from three independent experiments. Data were presented as Mean ± SEM. *p

    Article Snippet: Cells were then double-labeled with primary antibodies against TH (Santa Cruz; mouse monoclonal; 1:1000) and Nurr1 (Sigma; rabbit polyclonal; 1:1000) overnight at 4°C in a humidified chamber.

    Techniques: Inhibition, Multiple Displacement Amplification, Expressing, Transfection, Western Blot

    METH caused differential changes in Nurr1 and Pitx3 mRNA and protein levels in the midbrain. METH injections resulted in significant decreases in Nurr1 expression in all the groups (A). In contrast, METH challenges caused significant increases in Pitx3 mRNA expression only in the METH-preconditioned rats (B). Consistent with the mRNA data, toxic METH challenges caused decreases in Nurr1 protein levels in all the groups (C). In contrast, Pitx3 protein levels were increased by METH (D). The values represent means ± SEM in comparison to the saline-pretreated challenged with saline group (SSS). N = 3–6 animals per group. Keys to statistics are as described in the legend to Fig. 4 .

    Journal: PLoS ONE

    Article Title: Chronic Methamphetamine Administration Causes Differential Regulation of Transcription Factors in the Rat Midbrain

    doi: 10.1371/journal.pone.0019179

    Figure Lengend Snippet: METH caused differential changes in Nurr1 and Pitx3 mRNA and protein levels in the midbrain. METH injections resulted in significant decreases in Nurr1 expression in all the groups (A). In contrast, METH challenges caused significant increases in Pitx3 mRNA expression only in the METH-preconditioned rats (B). Consistent with the mRNA data, toxic METH challenges caused decreases in Nurr1 protein levels in all the groups (C). In contrast, Pitx3 protein levels were increased by METH (D). The values represent means ± SEM in comparison to the saline-pretreated challenged with saline group (SSS). N = 3–6 animals per group. Keys to statistics are as described in the legend to Fig. 4 .

    Article Snippet: Effects of METH on midbrain Nurr1 and Pitx3 mRNA and protein levels in the midbrain We measured the expression of NURR1 and PITX3 because they participate the molecular regulation of TH and DAT levels during both developmental stages and in adult life . presents the effects of METH on their expression in the midbrain.

    Techniques: Expressing

    Protein expression of synapsins, CREB, and Nurr1 in mouse primary cortical neurons following IC delivery of recombinant h-αSyn WT species. ( A ) IC delivery of exogenous fluorophore-conjugated antibodies using Chariot in primary cortical neurons 60 min postapplication. Neurons were labeled with MAP2 (magenta). (Scale bars: 10 μm.) ( B ) Representative Western blot (WB) images for αSyn in selected fractions following SEC segregation of recombinant monomers and multimers. Fraction #50 was enriched in monomers, whereas #42 was enriched in αSyn multimers. ( C ) Representative Western blot images for αSyn using lysates of cells subjected to IC delivery of fractions #50 and #42 (250 nM, monomer equivalent). ( D ) Representative Western blot analysis of synapsin expression in primary neurons 6 h after intraneuronal delivery of recombinant monomers or oligomers isolated by SEC. Actin was used as internal loading control. ( E ) Quantification of the relative expression levels of synapsins following intraneuronal delivery of recombinant h-αSyn WT species. (ANOVA followed by Student t test with Bonferroni correction, n = 4–6 dishes per group.) ( F ) Western blot analysis with 4D6 antibodies following Clear Native (CN)-PAGE segregation of SEC fraction #48 using IC lysates from TgI2.2, WT, or SNCA-null (KO) mice. Oligomeric recombinant h-αSyn WT (0.25 μg) was used as control. ( G ) Representative Western blot images documenting the protein abundance for pS133-CREB, total CREB, Nurr1, and the neuronal nuclear protein NeuN in membrane lysates from primary neurons treated with αSyn species delivered with Chariot. Please note the selective reductions in phosphorylated (p)CREB and Nurr1, whereas NeuN amounts remain unchanged. Veh., vehicle.

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

    Article Title: Selective lowering of synapsins induced by oligomeric α-synuclein exacerbates memory deficits

    doi: 10.1073/pnas.1704698114

    Figure Lengend Snippet: Protein expression of synapsins, CREB, and Nurr1 in mouse primary cortical neurons following IC delivery of recombinant h-αSyn WT species. ( A ) IC delivery of exogenous fluorophore-conjugated antibodies using Chariot in primary cortical neurons 60 min postapplication. Neurons were labeled with MAP2 (magenta). (Scale bars: 10 μm.) ( B ) Representative Western blot (WB) images for αSyn in selected fractions following SEC segregation of recombinant monomers and multimers. Fraction #50 was enriched in monomers, whereas #42 was enriched in αSyn multimers. ( C ) Representative Western blot images for αSyn using lysates of cells subjected to IC delivery of fractions #50 and #42 (250 nM, monomer equivalent). ( D ) Representative Western blot analysis of synapsin expression in primary neurons 6 h after intraneuronal delivery of recombinant monomers or oligomers isolated by SEC. Actin was used as internal loading control. ( E ) Quantification of the relative expression levels of synapsins following intraneuronal delivery of recombinant h-αSyn WT species. (ANOVA followed by Student t test with Bonferroni correction, n = 4–6 dishes per group.) ( F ) Western blot analysis with 4D6 antibodies following Clear Native (CN)-PAGE segregation of SEC fraction #48 using IC lysates from TgI2.2, WT, or SNCA-null (KO) mice. Oligomeric recombinant h-αSyn WT (0.25 μg) was used as control. ( G ) Representative Western blot images documenting the protein abundance for pS133-CREB, total CREB, Nurr1, and the neuronal nuclear protein NeuN in membrane lysates from primary neurons treated with αSyn species delivered with Chariot. Please note the selective reductions in phosphorylated (p)CREB and Nurr1, whereas NeuN amounts remain unchanged. Veh., vehicle.

    Article Snippet: The following primary antibodies were used in this study: LB509 (1:5,000–10,000), 4D6 (1:5,000), 4B12 (1:5,000), 6E10 (1:2,500; Covance and BioLegend), antisynapsin-I/II (1:1,000) (#106002), antisynapsin-III (1:1,000) (#106303), anticomplexin-1/2 (1:5,000; Synaptic Systems Inc), anti–β-Syn (1:1,000), anti-SYP (1:25,000), anti-NeuN (1:5,000), anti-CREB (1:2,000), and anti–pSer133-CREB (1:1,000; EMD Millipore), anti-Nurr1 (1:1,000; ThermoFisher Scientific and Santa Cruz Biotechnology), αSyn C-20 (1:1,000; Santa Cruz Biotechnology), rabbit-host antiactin (1:10,000; Sigma-Aldrich), mouse-host anti-actin (1:10,000; Pierce), Syn-33 (1:500), F8H7 (1:500), A11 (1:2,000), OC (1:2,000), and Officer (1:2,000; generated in-house by the R.K. laboratory).

    Techniques: Expressing, Recombinant, Labeling, Western Blot, Size-exclusion Chromatography, Isolation, Clear Native PAGE, Mouse Assay

    αSyn oligomers down-regulate SYN1 and SYN2 gene expression through CREB and Nurr1. ( A ) Age-dependent changes of SYN1 , SYN2 , CPLX1 , CPLX2 , SYP , and SYT1 gene expression by rt-qPCR analysis in the forebrain of TgI2.2 mice. Two-way ANOVA revealed a significant effect of transgene ( F = 37.18, P

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

    Article Title: Selective lowering of synapsins induced by oligomeric α-synuclein exacerbates memory deficits

    doi: 10.1073/pnas.1704698114

    Figure Lengend Snippet: αSyn oligomers down-regulate SYN1 and SYN2 gene expression through CREB and Nurr1. ( A ) Age-dependent changes of SYN1 , SYN2 , CPLX1 , CPLX2 , SYP , and SYT1 gene expression by rt-qPCR analysis in the forebrain of TgI2.2 mice. Two-way ANOVA revealed a significant effect of transgene ( F = 37.18, P

    Article Snippet: The following primary antibodies were used in this study: LB509 (1:5,000–10,000), 4D6 (1:5,000), 4B12 (1:5,000), 6E10 (1:2,500; Covance and BioLegend), antisynapsin-I/II (1:1,000) (#106002), antisynapsin-III (1:1,000) (#106303), anticomplexin-1/2 (1:5,000; Synaptic Systems Inc), anti–β-Syn (1:1,000), anti-SYP (1:25,000), anti-NeuN (1:5,000), anti-CREB (1:2,000), and anti–pSer133-CREB (1:1,000; EMD Millipore), anti-Nurr1 (1:1,000; ThermoFisher Scientific and Santa Cruz Biotechnology), αSyn C-20 (1:1,000; Santa Cruz Biotechnology), rabbit-host antiactin (1:10,000; Sigma-Aldrich), mouse-host anti-actin (1:10,000; Pierce), Syn-33 (1:500), F8H7 (1:500), A11 (1:2,000), OC (1:2,000), and Officer (1:2,000; generated in-house by the R.K. laboratory).

    Techniques: Expressing, Quantitative RT-PCR, Mouse Assay

    Western blot analysis of Nurr1 protein expression in hippocampi derived from young, adult and aged gerbils. The RODs of immunoblot bands are shown as percentage values. Data are presented as the means ± standard error of the mean. *P

    Journal: Biomedical Reports

    Article Title: Age-dependent decrease of Nurr1 protein expression in the gerbil hippocampus

    doi: 10.3892/br.2018.1094

    Figure Lengend Snippet: Western blot analysis of Nurr1 protein expression in hippocampi derived from young, adult and aged gerbils. The RODs of immunoblot bands are shown as percentage values. Data are presented as the means ± standard error of the mean. *P

    Article Snippet: Following three washes with PBS with Tween-20 (PBST; each for 5 min), the membranes were incubated with rabbit anti-Nurr1 (PA5-13416; 1:500; Invitrogen; Thermo Fisher Scientific, Inc.) overnight at 4°C.

    Techniques: Western Blot, Expressing, Derivative Assay

    Nurr1 immunohistochemistry in hippocampal regions of young, adult and aged gerbils. Nurr1 expression was detected in the (A-C) CA1 and (D-F) CA3 regions and (G-I) DG. In the young group, strong Nurr1 immunoreactivity was detected in pyramidal neurons of the SP in the CA1 and 3 regions (arrows) and in granule cells of the GCL in the DG (arrowheads). Nurr1 immunoreactivity in the SP and GCL gradually decreased in the adult and aged groups. Magnification, ×20; scale bar, 100 µm. Nurr1, nuclear receptor related-1 protein; DG, dentate gyrus; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum; ML, molecular layer; GCL, granule cell layer; PL, polymorp hic layer.

    Journal: Biomedical Reports

    Article Title: Age-dependent decrease of Nurr1 protein expression in the gerbil hippocampus

    doi: 10.3892/br.2018.1094

    Figure Lengend Snippet: Nurr1 immunohistochemistry in hippocampal regions of young, adult and aged gerbils. Nurr1 expression was detected in the (A-C) CA1 and (D-F) CA3 regions and (G-I) DG. In the young group, strong Nurr1 immunoreactivity was detected in pyramidal neurons of the SP in the CA1 and 3 regions (arrows) and in granule cells of the GCL in the DG (arrowheads). Nurr1 immunoreactivity in the SP and GCL gradually decreased in the adult and aged groups. Magnification, ×20; scale bar, 100 µm. Nurr1, nuclear receptor related-1 protein; DG, dentate gyrus; SO, stratum oriens; SP, stratum pyramidale; SR, stratum radiatum; ML, molecular layer; GCL, granule cell layer; PL, polymorp hic layer.

    Article Snippet: Following three washes with PBS with Tween-20 (PBST; each for 5 min), the membranes were incubated with rabbit anti-Nurr1 (PA5-13416; 1:500; Invitrogen; Thermo Fisher Scientific, Inc.) overnight at 4°C.

    Techniques: Immunohistochemistry, Expressing, Hydrophobic Interaction Chromatography

    Temporal changes of Aβ deposits and Nurr1-positive cell density in the subiculum area during disease progression. (A) Representative pictures of Nurr1 and 4G8 double-labeling in the subiculum of 5XFAD mice at 2, 4 and 6 months of ages. Scale bar=20 μm. (B) Representative figures of Nurr1-positive cells in the subiculum of 5XFAD mice. Scale bar=20 μm. (C) Quantification of Nurr1-positive cells in the subiculum of wild-type littermates and 5XFAD mice (n=4~5). *

    Journal: Journal of neurochemistry

    Article Title: Correlation between Orphan Nuclear Receptor Nurr1 expression and Amyloid deposition in 5XFAD mice, an animal model of Alzheimer’s disease

    doi: 10.1111/jnc.12935

    Figure Lengend Snippet: Temporal changes of Aβ deposits and Nurr1-positive cell density in the subiculum area during disease progression. (A) Representative pictures of Nurr1 and 4G8 double-labeling in the subiculum of 5XFAD mice at 2, 4 and 6 months of ages. Scale bar=20 μm. (B) Representative figures of Nurr1-positive cells in the subiculum of 5XFAD mice. Scale bar=20 μm. (C) Quantification of Nurr1-positive cells in the subiculum of wild-type littermates and 5XFAD mice (n=4~5). *

    Article Snippet: The anti-Nurr1 specific antibodies were obtained by passing rabbit sera through a Protein A Plus spin column (Thermo, 89978) followed by passage through an AminoLink column (Pierce cat# 20381) coupled with Nor1 LBD and then through an AminoLink column coupled with Nur77 LBD.

    Techniques: Labeling, Mouse Assay

    Immunoreactivity of Nurr1 antibody in the subiculum (A) and the frontal cortex (B) of wild-type mice. Nurr1-expressing cells were restricted to specific cortical layers and subiculum (indicated by white circle and box). (C and D) A comparison of Nurr1-expressed areas with amyloid-deposited regions in the brain of 5XFAD mice using 4G8 antibody and Nurr1 antibody. Nurr1-positive cells are detected in subiculum (C, indicated by dotted white circle) and specific cortical layers 5/6 (D) of 2 month-old 5XFAD mice. Remarkably, Nurr1 is highly expressed in two amyloid-deposited regions, deep cortical layers and subiculum of 5XFAD mice. (E) Representative high magnification images of Nurr1 and 4G8 double labeling in the subiculum and cerebral cortex of 2 months old 5XFAD mice. Scale bar=20 μm (A–C and E) and 100 μm (D).

    Journal: Journal of neurochemistry

    Article Title: Correlation between Orphan Nuclear Receptor Nurr1 expression and Amyloid deposition in 5XFAD mice, an animal model of Alzheimer’s disease

    doi: 10.1111/jnc.12935

    Figure Lengend Snippet: Immunoreactivity of Nurr1 antibody in the subiculum (A) and the frontal cortex (B) of wild-type mice. Nurr1-expressing cells were restricted to specific cortical layers and subiculum (indicated by white circle and box). (C and D) A comparison of Nurr1-expressed areas with amyloid-deposited regions in the brain of 5XFAD mice using 4G8 antibody and Nurr1 antibody. Nurr1-positive cells are detected in subiculum (C, indicated by dotted white circle) and specific cortical layers 5/6 (D) of 2 month-old 5XFAD mice. Remarkably, Nurr1 is highly expressed in two amyloid-deposited regions, deep cortical layers and subiculum of 5XFAD mice. (E) Representative high magnification images of Nurr1 and 4G8 double labeling in the subiculum and cerebral cortex of 2 months old 5XFAD mice. Scale bar=20 μm (A–C and E) and 100 μm (D).

    Article Snippet: The anti-Nurr1 specific antibodies were obtained by passing rabbit sera through a Protein A Plus spin column (Thermo, 89978) followed by passage through an AminoLink column (Pierce cat# 20381) coupled with Nor1 LBD and then through an AminoLink column coupled with Nur77 LBD.

    Techniques: Mouse Assay, Expressing, Labeling

    (A) Murine sequence comparison and homology of the DBD and LBD domains of three NR4A members. (B) Production and isolation of Nurr1, Nor1, and Nur77 LBDs for antibody production and purification. Lane 1, Nur77-LBD; lane 2, Nor1-LBD; lane 3, Nurr1-LBD. Each LBD with histidine tag (molecular weight of 26, 27, and 30 kDa for Nur77, Nor1, Nurr1, respectively) was expressed in E. coli and purified to > 90% purity by histidine tag purification. (C) Step-wise purification of Nurr1-specific antibody: Although our Nurr1 antibody exhibited strong binding to Nurr1 protein, it also showed cross-reactivity to Nor1 and Nur77 proteins. Following two consecutive pre-adsorptions against Nor1 and Nur77 LBDs, the final Nurr1 antibody did not show any detectable cross-reactivity to Nor1 and Nur77. Each lane contained 20 μg of CHO cell extracts expressing Nur77-LBD (lane 1), Nor1-LBD (lane 2), and Nurr1-LBD (lane 3). (D) Comparison of different Nurr1 antibodies cross-reactivities: While our purified #19 antibody was highly specific for Nurr1 LBD, two commercially available antibodies showed cross-reactivities to Nor1 and Nur77 LBDs. Each lane contained 20 μg of CHO cell extracts expressing Nur77-LBD (lane 1), Nor1-LBD (lane 2), and Nurr1-LBD (lane 3). (E) Immunoreactivity of Nurr1 in the hippocampus of C57BL/6 mice. Nurr1-positive cells are abundantly expressed in CA1 of the hippocampus. Hippocampal CA3 exhibited moderate numbers of Nurr1-immunoreactive cells. The granule cell layer (GCL) of the dentate gyrus and the CA2 region showed the weak Nurr1immunoreactivity. Scale bar=100 μm.

    Journal: Journal of neurochemistry

    Article Title: Correlation between Orphan Nuclear Receptor Nurr1 expression and Amyloid deposition in 5XFAD mice, an animal model of Alzheimer’s disease

    doi: 10.1111/jnc.12935

    Figure Lengend Snippet: (A) Murine sequence comparison and homology of the DBD and LBD domains of three NR4A members. (B) Production and isolation of Nurr1, Nor1, and Nur77 LBDs for antibody production and purification. Lane 1, Nur77-LBD; lane 2, Nor1-LBD; lane 3, Nurr1-LBD. Each LBD with histidine tag (molecular weight of 26, 27, and 30 kDa for Nur77, Nor1, Nurr1, respectively) was expressed in E. coli and purified to > 90% purity by histidine tag purification. (C) Step-wise purification of Nurr1-specific antibody: Although our Nurr1 antibody exhibited strong binding to Nurr1 protein, it also showed cross-reactivity to Nor1 and Nur77 proteins. Following two consecutive pre-adsorptions against Nor1 and Nur77 LBDs, the final Nurr1 antibody did not show any detectable cross-reactivity to Nor1 and Nur77. Each lane contained 20 μg of CHO cell extracts expressing Nur77-LBD (lane 1), Nor1-LBD (lane 2), and Nurr1-LBD (lane 3). (D) Comparison of different Nurr1 antibodies cross-reactivities: While our purified #19 antibody was highly specific for Nurr1 LBD, two commercially available antibodies showed cross-reactivities to Nor1 and Nur77 LBDs. Each lane contained 20 μg of CHO cell extracts expressing Nur77-LBD (lane 1), Nor1-LBD (lane 2), and Nurr1-LBD (lane 3). (E) Immunoreactivity of Nurr1 in the hippocampus of C57BL/6 mice. Nurr1-positive cells are abundantly expressed in CA1 of the hippocampus. Hippocampal CA3 exhibited moderate numbers of Nurr1-immunoreactive cells. The granule cell layer (GCL) of the dentate gyrus and the CA2 region showed the weak Nurr1immunoreactivity. Scale bar=100 μm.

    Article Snippet: The anti-Nurr1 specific antibodies were obtained by passing rabbit sera through a Protein A Plus spin column (Thermo, 89978) followed by passage through an AminoLink column (Pierce cat# 20381) coupled with Nor1 LBD and then through an AminoLink column coupled with Nur77 LBD.

    Techniques: Sequencing, Isolation, Purification, Molecular Weight, Binding Assay, Expressing, Mouse Assay

    Nurr1 expression in the ventral midbrain of 5XFAD mice. (A) The distribution pattern of 4G8-stained amyloid plaques in the coronal brain section of 4 month-old 5XFAD mice. The ventral midbrain, in which dopaminergic substantia nigra neurons and ventral tegmental area neurons are located, remains devoid of amyloid deposits in 5XFAD mice. (B) Nurr1-immunoreactive cells in substantia nigra of wild-type (WT) and 5XFAD mice. There was no reduction of Nurr1-expressing cells in ventral midbrain of 5XFAD mice at 4 months of age, compared with wild-type littermates. Scale bar=50 μm.

    Journal: Journal of neurochemistry

    Article Title: Correlation between Orphan Nuclear Receptor Nurr1 expression and Amyloid deposition in 5XFAD mice, an animal model of Alzheimer’s disease

    doi: 10.1111/jnc.12935

    Figure Lengend Snippet: Nurr1 expression in the ventral midbrain of 5XFAD mice. (A) The distribution pattern of 4G8-stained amyloid plaques in the coronal brain section of 4 month-old 5XFAD mice. The ventral midbrain, in which dopaminergic substantia nigra neurons and ventral tegmental area neurons are located, remains devoid of amyloid deposits in 5XFAD mice. (B) Nurr1-immunoreactive cells in substantia nigra of wild-type (WT) and 5XFAD mice. There was no reduction of Nurr1-expressing cells in ventral midbrain of 5XFAD mice at 4 months of age, compared with wild-type littermates. Scale bar=50 μm.

    Article Snippet: The anti-Nurr1 specific antibodies were obtained by passing rabbit sera through a Protein A Plus spin column (Thermo, 89978) followed by passage through an AminoLink column (Pierce cat# 20381) coupled with Nor1 LBD and then through an AminoLink column coupled with Nur77 LBD.

    Techniques: Expressing, Mouse Assay, Staining

    Effects of intrastriatal injection of Lv-Nurr1-MSCs on apomorphine-induced rotation asymmetry in rats after 6-hydroxydopamine (6-OHDA) lesion. Average rotational turns/min are showed at different time points (1, 2 and 4 weeks, n = 5 for each group). The average rotational rates of PD rats in the Lv-MSCs group (10.57 ± 1.86 at 1 week, 7.64 ± 1.56 at 2 weeks and 6.71 ± 1.87 at 4 weeks) significantly decreased ( # P

    Journal: American Journal of Translational Research

    Article Title: The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’s disease

    doi:

    Figure Lengend Snippet: Effects of intrastriatal injection of Lv-Nurr1-MSCs on apomorphine-induced rotation asymmetry in rats after 6-hydroxydopamine (6-OHDA) lesion. Average rotational turns/min are showed at different time points (1, 2 and 4 weeks, n = 5 for each group). The average rotational rates of PD rats in the Lv-MSCs group (10.57 ± 1.86 at 1 week, 7.64 ± 1.56 at 2 weeks and 6.71 ± 1.87 at 4 weeks) significantly decreased ( # P

    Article Snippet: Western blot was performed to assess whether Nurr1 was expressed and secreted by MSCs infected with GV287-Nurr1 virus and to examine the expression changes of Nurr1, TH and DAT in SN tissue after the Nurr1 gene-modified MSCs transplantation.

    Techniques: Injection

    Effects of Lv-Nurr1-MSCs on the number of nigral TH-positive neurons, the density of striatal TH-positive fibers, and the expression of Nurr1, TH and DAT in mRNA and protein level of the substantia nigra. The number of nigral TH-positive neurons and the density of striatal TH-positive fibers in the group of Lv-Nurr1-MSCs-injected were more than other group. TH-positive cells in the SN (A1-A4); TH-positive fiber in the striatum (The data has been shown in the text) (B1-B4); Representative RT-PCR of Nurr1, TH and DAT in the SN. GAPDH was used as an internal control (C). The relative mRNA level of Nurr1, TH and DAT was analyzed by RT-PCR (* P

    Journal: American Journal of Translational Research

    Article Title: The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’s disease

    doi:

    Figure Lengend Snippet: Effects of Lv-Nurr1-MSCs on the number of nigral TH-positive neurons, the density of striatal TH-positive fibers, and the expression of Nurr1, TH and DAT in mRNA and protein level of the substantia nigra. The number of nigral TH-positive neurons and the density of striatal TH-positive fibers in the group of Lv-Nurr1-MSCs-injected were more than other group. TH-positive cells in the SN (A1-A4); TH-positive fiber in the striatum (The data has been shown in the text) (B1-B4); Representative RT-PCR of Nurr1, TH and DAT in the SN. GAPDH was used as an internal control (C). The relative mRNA level of Nurr1, TH and DAT was analyzed by RT-PCR (* P

    Article Snippet: Western blot was performed to assess whether Nurr1 was expressed and secreted by MSCs infected with GV287-Nurr1 virus and to examine the expression changes of Nurr1, TH and DAT in SN tissue after the Nurr1 gene-modified MSCs transplantation.

    Techniques: Expressing, Injection, Reverse Transcription Polymerase Chain Reaction

    Effects of intrastriatal injection of Lv-Nurr1-MSCs on the morphology of the CD11b-, GFAP-, COX-2- and TNF-α-positive cells and the integrated absorbance (IA) of CD11b, GFAP, COX-2 and TNF-α in mRNA level of the substantia nigra. CD11b-positive cells (A1-A4); GFAP-positive cells (B1-B4); COX-2-positive cells (C1-C4); TNF-α-positive cells (D1-D4); Representative RT-PCR of CD11b, GFAP, COX-2 and TNF-α in the SN. GAPDH was used as an internal control (E). The relative mRNA level of CD11b, GFAP, COX-2 and TNF-α was analyzed by RT-PCR ( Δ P

    Journal: American Journal of Translational Research

    Article Title: The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’s disease

    doi:

    Figure Lengend Snippet: Effects of intrastriatal injection of Lv-Nurr1-MSCs on the morphology of the CD11b-, GFAP-, COX-2- and TNF-α-positive cells and the integrated absorbance (IA) of CD11b, GFAP, COX-2 and TNF-α in mRNA level of the substantia nigra. CD11b-positive cells (A1-A4); GFAP-positive cells (B1-B4); COX-2-positive cells (C1-C4); TNF-α-positive cells (D1-D4); Representative RT-PCR of CD11b, GFAP, COX-2 and TNF-α in the SN. GAPDH was used as an internal control (E). The relative mRNA level of CD11b, GFAP, COX-2 and TNF-α was analyzed by RT-PCR ( Δ P

    Article Snippet: Western blot was performed to assess whether Nurr1 was expressed and secreted by MSCs infected with GV287-Nurr1 virus and to examine the expression changes of Nurr1, TH and DAT in SN tissue after the Nurr1 gene-modified MSCs transplantation.

    Techniques: Injection, IA, Reverse Transcription Polymerase Chain Reaction

    Survival and migration and differentiation of implanted cells in the rat brain of the Lv-Nurr1-MSCs group four weeks after transplantation by immunohistochemical and double immunofluorescence staining. Lv-Nurr1-MSCs were injected into the striatum with microsyringe. The number of Nurr1-positive cells in the right striatum was significantly more than that in the left side, and cell migration was visible. Nurr1-positive cells were in the different part of the brain. (A). Nurr1-positive cells were fusiform or polygonal, and some like neurons. (B) is the enlarged view of (A) (B). Double immunofluorescence staining of TH (red) and Nurr1 (green) and nucleus (blue) (C-E). The number of rats in LV-Nurr1-MSCs group was five. Scale bar: (A) = 500 μm; (B) = 50 μm; (C-E) = 20 μm.

    Journal: American Journal of Translational Research

    Article Title: The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’s disease

    doi:

    Figure Lengend Snippet: Survival and migration and differentiation of implanted cells in the rat brain of the Lv-Nurr1-MSCs group four weeks after transplantation by immunohistochemical and double immunofluorescence staining. Lv-Nurr1-MSCs were injected into the striatum with microsyringe. The number of Nurr1-positive cells in the right striatum was significantly more than that in the left side, and cell migration was visible. Nurr1-positive cells were in the different part of the brain. (A). Nurr1-positive cells were fusiform or polygonal, and some like neurons. (B) is the enlarged view of (A) (B). Double immunofluorescence staining of TH (red) and Nurr1 (green) and nucleus (blue) (C-E). The number of rats in LV-Nurr1-MSCs group was five. Scale bar: (A) = 500 μm; (B) = 50 μm; (C-E) = 20 μm.

    Article Snippet: Western blot was performed to assess whether Nurr1 was expressed and secreted by MSCs infected with GV287-Nurr1 virus and to examine the expression changes of Nurr1, TH and DAT in SN tissue after the Nurr1 gene-modified MSCs transplantation.

    Techniques: Migration, Transplantation Assay, Immunohistochemistry, Double Immunofluorescence Staining, Injection

    Expression of Nurr1, TH and DAT in the substantia nigra of PD rats detected by immunohistochemistry, RT-PCR and Western blot. Microphotographs showed the Nurr1, TH and DAT immunoreactive neurons (A). Representative RT-PCR of Nurr1, TH and DAT in normal and PD rats. GAPDH was used as an internal control (B). The relative mRNA level of Nurr1, TH and DAT was analyzed by RT-PCR (C). Representative Western blot of Nurr1, TH and DAT protein in normal and PD rats. β-actin was used as an internal control (D). The relative intensity of Nurr1, TH and DAT protein was analyzed by Western blot (E). Data are presented as mean ± SEM. * P

    Journal: American Journal of Translational Research

    Article Title: The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’s disease

    doi:

    Figure Lengend Snippet: Expression of Nurr1, TH and DAT in the substantia nigra of PD rats detected by immunohistochemistry, RT-PCR and Western blot. Microphotographs showed the Nurr1, TH and DAT immunoreactive neurons (A). Representative RT-PCR of Nurr1, TH and DAT in normal and PD rats. GAPDH was used as an internal control (B). The relative mRNA level of Nurr1, TH and DAT was analyzed by RT-PCR (C). Representative Western blot of Nurr1, TH and DAT protein in normal and PD rats. β-actin was used as an internal control (D). The relative intensity of Nurr1, TH and DAT protein was analyzed by Western blot (E). Data are presented as mean ± SEM. * P

    Article Snippet: Western blot was performed to assess whether Nurr1 was expressed and secreted by MSCs infected with GV287-Nurr1 virus and to examine the expression changes of Nurr1, TH and DAT in SN tissue after the Nurr1 gene-modified MSCs transplantation.

    Techniques: Expressing, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Western Blot

    Expression of Nurr1 gene in transduced rat MSCs. The morphological features of MSCs infected by lentivirus GV287-Nurr1 (MOI = 50) at different times (A-C) and MSCs transduced by lentivirus GV287 (D). MSCs infected by lentivirus GV287-Nurr1 after 24 h (A); MSCs infected by lentivirus GV287-Nurr1 after 48 h (B); MSCs infected by lentivirus GV287-Nurr1 after 72 h, and differentiated into neuron-like cells with monopolar or multipolar processes (C); MSCs infected by lentivirus GV287 after 72 h (D). Representative Western blot of Nurr1 protein in cell extracts and supernatant from MSCs after transduction with or without GV287-Nurr1. β-actin was used as an internal control (E). The relative intensity of Nurr1 protein was analyzed by Western blot (F). Data are presented as mean ± SEM. Scale bar: (A) = 100 μm; (B) = 50 μm; (C, D) = 20 μm.

    Journal: American Journal of Translational Research

    Article Title: The lentiviral-mediated Nurr1 genetic engineering mesenchymal stem cells protect dopaminergic neurons in a rat model of Parkinson’s disease

    doi:

    Figure Lengend Snippet: Expression of Nurr1 gene in transduced rat MSCs. The morphological features of MSCs infected by lentivirus GV287-Nurr1 (MOI = 50) at different times (A-C) and MSCs transduced by lentivirus GV287 (D). MSCs infected by lentivirus GV287-Nurr1 after 24 h (A); MSCs infected by lentivirus GV287-Nurr1 after 48 h (B); MSCs infected by lentivirus GV287-Nurr1 after 72 h, and differentiated into neuron-like cells with monopolar or multipolar processes (C); MSCs infected by lentivirus GV287 after 72 h (D). Representative Western blot of Nurr1 protein in cell extracts and supernatant from MSCs after transduction with or without GV287-Nurr1. β-actin was used as an internal control (E). The relative intensity of Nurr1 protein was analyzed by Western blot (F). Data are presented as mean ± SEM. Scale bar: (A) = 100 μm; (B) = 50 μm; (C, D) = 20 μm.

    Article Snippet: Western blot was performed to assess whether Nurr1 was expressed and secreted by MSCs infected with GV287-Nurr1 virus and to examine the expression changes of Nurr1, TH and DAT in SN tissue after the Nurr1 gene-modified MSCs transplantation.

    Techniques: Expressing, Infection, Western Blot, Transduction