15d-pgj 2 (cat Search Results


15d-pgj 2 (cat# 18570)  (Cayman Chemical)
90
Cayman Chemical 15d-pgj 2 (cat# 18570)
A ) Schematic shows upstream and downstream kinases in the AKT signaling network. B ) BMDMs were primed, or not, with LPS and then stimulated with oxPAPC (100, 50 or 25 μg ml −1 ). Phosphorylation of upstream AKT regulators (TBK1, IKKε, PDK1, PTEN and PI3K) was analyzed by immunoblot. Images are representative of three independent experiments. C ) BMDMs were primed, or not, with LPS (1 μg ml −1 ) and then stimulated with oxPAPC (100, 50 or 25 μg ml −1 ). The phosphorylation of the indicated mTORC1/2-associated proteins was analyzed after 1h by immunoblotting. Data are representative of three independent experiments. D ) WT and Nfe2l2 −/− BMDMs were primed, or not, with LPS (1 μg ml −1 ) and treated or not with oxPAPC (100 μg ml −1 ) for 1h. The indicated transcripts were analyzed by qPCR. n = 3, data are representative of three independent experiments and show means ± SEM. Statistical significance was calculated using two-way ANOVA and Sidak’s multiple comparisons test. E-F ) BMDMs were primed, or not, with LPS (1 μg ml −1 ) and treated or not with <t>15d-PGJ</t> 2 (20, 10 or 1 μM in E and 10 μM in F ) for 1h ( E ) or 24h ( F ). AKT phosphorylation and NRF2 accumulation were assessed by immunoblot ( E ). Data are representative of three independent experiments. IL-10 and TNF release was quantified by ELISA ( F ). n = 6, graphs are representative of three independent experiments and show means ± SEM. Statistical multiple comparisons were calculated by two-way ANOVA and Sidak’s test. G ) Expression of Akt isoforms (RNA-seq, Immgen.org database) in the indicated murine macrophage populations. H ) Normalized expression of Akt1 , Akt2 and Akt3 in BMDMs, analyzed by qPCR. n = 3, data are representative of three independent experiments and show means ± SEM. I, J ) Microscale thermophoresis (MST) analysis of oxPAPC interactions with AKT2 ( I ) and AKT3 ( J ). Traces of fluorescently labeled human recombinant AKT incubated with oxPAPC (1000, 500, 250,125, 62.5, 31.25, 15.62, 7.8, 3.9, 1.9, 0.97, 0.488, 0.24, 0.12, 0.0061 and 0.030 μM) or DPPC (500, 250,125, 62.5, 31.25, 15.62, 7.8, 3.9, 1.9, 0.97, 0.488, 0.24, 0.12, 0.0061 and 0.030 μM). oxPAPC-AKT binding curve was derived from the quantification of normalized fluorescence changes. n = 3, graph shows means ± SD. Images are representative of three independent experiments. K, L ) Active human recombinant AKT2 ( K ) or AKT3 ( L ) were incubated with oxPAPC and a kinase-specific FRET-peptide substrate (Z-’LYTE). Kinase inhibition was measured as the ability of lipid to block substrate phosphorylation. n = 4, graph shows means ± SD. Data are representative of three independent experiments.
15d Pgj 2 (Cat# 18570), supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/15d-pgj 2 (cat# 18570)/product/Cayman Chemical
Average 90 stars, based on 1 article reviews
15d-pgj 2 (cat# 18570) - by Bioz Stars, 2026-06
90/100 stars
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15d-pgj 2 antibody  (Enzo Biochem)
90
Enzo Biochem 15d-pgj 2 antibody
<t>15d-PGJ</t> 2 induced NRROS expression in RBA-1 cells. a , b RBA-1 cells were treated with 1, 5, or 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of a NRROS protein and b mRNA expression were analyzed by Western blot and RT/real-time PCR, respectively. Lower panel of a indicates the quantitation of 10 μM 15d-PGJ 2 treatment. c RBA-1 cells were pretreated with actinomycin D (ActD, 1 μM), mithramycin A (Mith A, 1 μM), GW9662 (GW, 10 nM), LY294002 (10 μM), Akti VIII (1 μM), or AS1842856 (AS, 0.3 μM) for 1 h and then treated with 10 μM 15d-PGJ 2 for 2 h. The levels of NRROS mRNA expression were analyzed by RT/real-time PCR. The data are expressed as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significant difference as indicated
15d Pgj 2 Antibody, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/15d-pgj 2 antibody/product/Enzo Biochem
Average 90 stars, based on 1 article reviews
15d-pgj 2 antibody - by Bioz Stars, 2026-06
90/100 stars
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15d pgj2  (Santa Cruz Biotechnology)
90
Santa Cruz Biotechnology 15d pgj2
Comparison of neurological deficits score.
15d Pgj2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/15d pgj2/product/Santa Cruz Biotechnology
Average 90 stars, based on 1 article reviews
15d pgj2 - by Bioz Stars, 2026-06
90/100 stars
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Image Search Results


A ) Schematic shows upstream and downstream kinases in the AKT signaling network. B ) BMDMs were primed, or not, with LPS and then stimulated with oxPAPC (100, 50 or 25 μg ml −1 ). Phosphorylation of upstream AKT regulators (TBK1, IKKε, PDK1, PTEN and PI3K) was analyzed by immunoblot. Images are representative of three independent experiments. C ) BMDMs were primed, or not, with LPS (1 μg ml −1 ) and then stimulated with oxPAPC (100, 50 or 25 μg ml −1 ). The phosphorylation of the indicated mTORC1/2-associated proteins was analyzed after 1h by immunoblotting. Data are representative of three independent experiments. D ) WT and Nfe2l2 −/− BMDMs were primed, or not, with LPS (1 μg ml −1 ) and treated or not with oxPAPC (100 μg ml −1 ) for 1h. The indicated transcripts were analyzed by qPCR. n = 3, data are representative of three independent experiments and show means ± SEM. Statistical significance was calculated using two-way ANOVA and Sidak’s multiple comparisons test. E-F ) BMDMs were primed, or not, with LPS (1 μg ml −1 ) and treated or not with 15d-PGJ 2 (20, 10 or 1 μM in E and 10 μM in F ) for 1h ( E ) or 24h ( F ). AKT phosphorylation and NRF2 accumulation were assessed by immunoblot ( E ). Data are representative of three independent experiments. IL-10 and TNF release was quantified by ELISA ( F ). n = 6, graphs are representative of three independent experiments and show means ± SEM. Statistical multiple comparisons were calculated by two-way ANOVA and Sidak’s test. G ) Expression of Akt isoforms (RNA-seq, Immgen.org database) in the indicated murine macrophage populations. H ) Normalized expression of Akt1 , Akt2 and Akt3 in BMDMs, analyzed by qPCR. n = 3, data are representative of three independent experiments and show means ± SEM. I, J ) Microscale thermophoresis (MST) analysis of oxPAPC interactions with AKT2 ( I ) and AKT3 ( J ). Traces of fluorescently labeled human recombinant AKT incubated with oxPAPC (1000, 500, 250,125, 62.5, 31.25, 15.62, 7.8, 3.9, 1.9, 0.97, 0.488, 0.24, 0.12, 0.0061 and 0.030 μM) or DPPC (500, 250,125, 62.5, 31.25, 15.62, 7.8, 3.9, 1.9, 0.97, 0.488, 0.24, 0.12, 0.0061 and 0.030 μM). oxPAPC-AKT binding curve was derived from the quantification of normalized fluorescence changes. n = 3, graph shows means ± SD. Images are representative of three independent experiments. K, L ) Active human recombinant AKT2 ( K ) or AKT3 ( L ) were incubated with oxPAPC and a kinase-specific FRET-peptide substrate (Z-’LYTE). Kinase inhibition was measured as the ability of lipid to block substrate phosphorylation. n = 4, graph shows means ± SD. Data are representative of three independent experiments.

Journal: bioRxiv

Article Title: Host-derived oxidized phospholipids initiate effector-triggered immunity fostering lethality upon microbial encounter

doi: 10.1101/2023.11.21.568047

Figure Lengend Snippet: A ) Schematic shows upstream and downstream kinases in the AKT signaling network. B ) BMDMs were primed, or not, with LPS and then stimulated with oxPAPC (100, 50 or 25 μg ml −1 ). Phosphorylation of upstream AKT regulators (TBK1, IKKε, PDK1, PTEN and PI3K) was analyzed by immunoblot. Images are representative of three independent experiments. C ) BMDMs were primed, or not, with LPS (1 μg ml −1 ) and then stimulated with oxPAPC (100, 50 or 25 μg ml −1 ). The phosphorylation of the indicated mTORC1/2-associated proteins was analyzed after 1h by immunoblotting. Data are representative of three independent experiments. D ) WT and Nfe2l2 −/− BMDMs were primed, or not, with LPS (1 μg ml −1 ) and treated or not with oxPAPC (100 μg ml −1 ) for 1h. The indicated transcripts were analyzed by qPCR. n = 3, data are representative of three independent experiments and show means ± SEM. Statistical significance was calculated using two-way ANOVA and Sidak’s multiple comparisons test. E-F ) BMDMs were primed, or not, with LPS (1 μg ml −1 ) and treated or not with 15d-PGJ 2 (20, 10 or 1 μM in E and 10 μM in F ) for 1h ( E ) or 24h ( F ). AKT phosphorylation and NRF2 accumulation were assessed by immunoblot ( E ). Data are representative of three independent experiments. IL-10 and TNF release was quantified by ELISA ( F ). n = 6, graphs are representative of three independent experiments and show means ± SEM. Statistical multiple comparisons were calculated by two-way ANOVA and Sidak’s test. G ) Expression of Akt isoforms (RNA-seq, Immgen.org database) in the indicated murine macrophage populations. H ) Normalized expression of Akt1 , Akt2 and Akt3 in BMDMs, analyzed by qPCR. n = 3, data are representative of three independent experiments and show means ± SEM. I, J ) Microscale thermophoresis (MST) analysis of oxPAPC interactions with AKT2 ( I ) and AKT3 ( J ). Traces of fluorescently labeled human recombinant AKT incubated with oxPAPC (1000, 500, 250,125, 62.5, 31.25, 15.62, 7.8, 3.9, 1.9, 0.97, 0.488, 0.24, 0.12, 0.0061 and 0.030 μM) or DPPC (500, 250,125, 62.5, 31.25, 15.62, 7.8, 3.9, 1.9, 0.97, 0.488, 0.24, 0.12, 0.0061 and 0.030 μM). oxPAPC-AKT binding curve was derived from the quantification of normalized fluorescence changes. n = 3, graph shows means ± SD. Images are representative of three independent experiments. K, L ) Active human recombinant AKT2 ( K ) or AKT3 ( L ) were incubated with oxPAPC and a kinase-specific FRET-peptide substrate (Z-’LYTE). Kinase inhibition was measured as the ability of lipid to block substrate phosphorylation. n = 4, graph shows means ± SD. Data are representative of three independent experiments.

Article Snippet: 15d-PGJ 2 (Cat# 18570) and 2-deoxy-D-Glucose (2-DG, Cat# 14325) were purchased from Cayman Chemicals.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, RNA Sequencing Assay, Microscale Thermophoresis, Labeling, Recombinant, Incubation, Binding Assay, Derivative Assay, Fluorescence, Inhibition, Blocking Assay

15d-PGJ 2 induced NRROS expression in RBA-1 cells. a , b RBA-1 cells were treated with 1, 5, or 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of a NRROS protein and b mRNA expression were analyzed by Western blot and RT/real-time PCR, respectively. Lower panel of a indicates the quantitation of 10 μM 15d-PGJ 2 treatment. c RBA-1 cells were pretreated with actinomycin D (ActD, 1 μM), mithramycin A (Mith A, 1 μM), GW9662 (GW, 10 nM), LY294002 (10 μM), Akti VIII (1 μM), or AS1842856 (AS, 0.3 μM) for 1 h and then treated with 10 μM 15d-PGJ 2 for 2 h. The levels of NRROS mRNA expression were analyzed by RT/real-time PCR. The data are expressed as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significant difference as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: 15d-PGJ 2 induced NRROS expression in RBA-1 cells. a , b RBA-1 cells were treated with 1, 5, or 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of a NRROS protein and b mRNA expression were analyzed by Western blot and RT/real-time PCR, respectively. Lower panel of a indicates the quantitation of 10 μM 15d-PGJ 2 treatment. c RBA-1 cells were pretreated with actinomycin D (ActD, 1 μM), mithramycin A (Mith A, 1 μM), GW9662 (GW, 10 nM), LY294002 (10 μM), Akti VIII (1 μM), or AS1842856 (AS, 0.3 μM) for 1 h and then treated with 10 μM 15d-PGJ 2 for 2 h. The levels of NRROS mRNA expression were analyzed by RT/real-time PCR. The data are expressed as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significant difference as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Quantitation Assay, Cell Culture

Analysis of rat NRTROS promoter and screening for 15d-PGJ 2 response elements. a The DNA sequences of rat NRROS promoter and transcription factor binding sites. The exon 1 regions are shown in boldface and (+ 1) indicates the first nucleotide of the mRNA. b RBA-1 cells were transfected with either rat (r)NRROS or (h)NRROS plasmid, and pCMV-β-gal DNA for 24 h, and then treated with 10 μM 15d-PGJ 2 for the indicated time intervals. The promoter activity was determined in the cell lysates using a promoter assay kit. c Scheme of the rat NRROS promoter region and reporter constructs. The position of deletion mutant, transcription initiation (+ 1), and location of exon 1 are indicated. Figures were not drawn to scale. d RBA-1 cells transfected with various NRROS constructs were treated with 10 μM 15d-PGJ 2 for 1 h. The pGL3B-Luc without the promoter was used as a control (Basic). Deletion mutants were used to analyze the essential promoter regions and 15d-PGJ 2 response element. RLU indicates the related luciferase units. Statistical analysis was determined using two-tailed unpaired Student’s t test. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: Analysis of rat NRTROS promoter and screening for 15d-PGJ 2 response elements. a The DNA sequences of rat NRROS promoter and transcription factor binding sites. The exon 1 regions are shown in boldface and (+ 1) indicates the first nucleotide of the mRNA. b RBA-1 cells were transfected with either rat (r)NRROS or (h)NRROS plasmid, and pCMV-β-gal DNA for 24 h, and then treated with 10 μM 15d-PGJ 2 for the indicated time intervals. The promoter activity was determined in the cell lysates using a promoter assay kit. c Scheme of the rat NRROS promoter region and reporter constructs. The position of deletion mutant, transcription initiation (+ 1), and location of exon 1 are indicated. Figures were not drawn to scale. d RBA-1 cells transfected with various NRROS constructs were treated with 10 μM 15d-PGJ 2 for 1 h. The pGL3B-Luc without the promoter was used as a control (Basic). Deletion mutants were used to analyze the essential promoter regions and 15d-PGJ 2 response element. RLU indicates the related luciferase units. Statistical analysis was determined using two-tailed unpaired Student’s t test. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Binding Assay, Transfection, Plasmid Preparation, Activity Assay, Promoter Assay, Construct, Mutagenesis, Control, Luciferase, Two Tailed Test, Cell Culture

15d-PGJ 2 -induced NRROS expression is mediated through PI3K, AKT, and FoxO1. RBA-1 cells were pretreated with different concentrations of a , d LY294002, b , e AKTi VIII, or c , f AS1842856 for 1 h and then incubated with 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of NRROS, phospho-AKT, phospho-FoxO1, AKT, FoxO1, and GAPDH protein were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: 15d-PGJ 2 -induced NRROS expression is mediated through PI3K, AKT, and FoxO1. RBA-1 cells were pretreated with different concentrations of a , d LY294002, b , e AKTi VIII, or c , f AS1842856 for 1 h and then incubated with 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of NRROS, phospho-AKT, phospho-FoxO1, AKT, FoxO1, and GAPDH protein were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Expressing, Incubation, Western Blot, Cell Culture

The roles of PI3K, AKT, and FoxO1 in 15d-PGJ 2 -induced NRROS expression are confirmed by transfection with respective siRNAs. RBA-1 cells were transfected with siRNA ( a , d p85; b , e AKT, or c , f FoxO1) or scramble siRNA for 48 h and then incubated with 10 μM 15d-PGJ 2 for 4 h ( a – c ) and for the indicated time intervals ( d – f ). The levels of NRROS, phospho-AKT, phospho-FoxO1, AKT, FoxO1, and GAPDH protein were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: The roles of PI3K, AKT, and FoxO1 in 15d-PGJ 2 -induced NRROS expression are confirmed by transfection with respective siRNAs. RBA-1 cells were transfected with siRNA ( a , d p85; b , e AKT, or c , f FoxO1) or scramble siRNA for 48 h and then incubated with 10 μM 15d-PGJ 2 for 4 h ( a – c ) and for the indicated time intervals ( d – f ). The levels of NRROS, phospho-AKT, phospho-FoxO1, AKT, FoxO1, and GAPDH protein were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Expressing, Transfection, Incubation, Western Blot, Cell Culture

15d-PGJ 2 stimulates the interaction between Sp1 and FoxO1 leading to NRROS expression. a RBA-1 cells were transfected with Sp1 siRNA and then incubated with 10 μM 15d-PGJ 2 for 4 h. The levels of NRROS, Sp1, and GAPDH were analyzed by Western blotting. b Schematic representation of rat NRROS promoter. Locations of primers used for ChIP assays (Sp1 and FoxO1) and transcription factors associated regions are indicated. c , d ChIP assays were performed using an anti-Sp1 or anti-FoxO1 antibody and then amplified the DNA fragments with Sp1 and FoxO1 primers, respectively. e , f RBA-1 cells were pretreated without or with mithramycin A or AS1842856 for 1 h and then incubated with 15d-PGJ 2 for 1 h. The association between FoxO1 and Sp1 on promoter was determined by ChIP assays. g Cells were treated with 10 μM 15d-PGJ 2 for the indicated time intervals. ChIP assays were performed using an anti-FoxO1 or anti-phospho-FoxO1 antibody. h Cells were treated with 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of protein expression in either cytoplasmic or nuclear fractions were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: 15d-PGJ 2 stimulates the interaction between Sp1 and FoxO1 leading to NRROS expression. a RBA-1 cells were transfected with Sp1 siRNA and then incubated with 10 μM 15d-PGJ 2 for 4 h. The levels of NRROS, Sp1, and GAPDH were analyzed by Western blotting. b Schematic representation of rat NRROS promoter. Locations of primers used for ChIP assays (Sp1 and FoxO1) and transcription factors associated regions are indicated. c , d ChIP assays were performed using an anti-Sp1 or anti-FoxO1 antibody and then amplified the DNA fragments with Sp1 and FoxO1 primers, respectively. e , f RBA-1 cells were pretreated without or with mithramycin A or AS1842856 for 1 h and then incubated with 15d-PGJ 2 for 1 h. The association between FoxO1 and Sp1 on promoter was determined by ChIP assays. g Cells were treated with 10 μM 15d-PGJ 2 for the indicated time intervals. ChIP assays were performed using an anti-FoxO1 or anti-phospho-FoxO1 antibody. h Cells were treated with 10 μM 15d-PGJ 2 for the indicated time intervals. The levels of protein expression in either cytoplasmic or nuclear fractions were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Expressing, Transfection, Incubation, Western Blot, Amplification, Cell Culture

NRROS expression via Nrf2 activation by 15d-PGJ 2 in RBA-1 cells. The cells were transfected with Nrf2 siRNA or scramble siRNA ( a , b ) for 48 h and then incubated with 10 μM 15d-PGJ 2 for 4 h or 6 h ( a ), and indicated time intervals ( b ). c The cells were pretreated with various concentration of anti-15d-PGJ2 antibody for 1 h and then incubated with 10 μM 15d-PGJ 2 for 6 h. The levels of NRROS, phospho-Nrf2, Nrf2, β-actin, and GAPDH protein were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: NRROS expression via Nrf2 activation by 15d-PGJ 2 in RBA-1 cells. The cells were transfected with Nrf2 siRNA or scramble siRNA ( a , b ) for 48 h and then incubated with 10 μM 15d-PGJ 2 for 4 h or 6 h ( a ), and indicated time intervals ( b ). c The cells were pretreated with various concentration of anti-15d-PGJ2 antibody for 1 h and then incubated with 10 μM 15d-PGJ 2 for 6 h. The levels of NRROS, phospho-Nrf2, Nrf2, β-actin, and GAPDH protein were analyzed by Western blotting. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). # p < 0.05, as compared with the respective values significantly different as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Expressing, Activation Assay, Transfection, Incubation, Concentration Assay, Western Blot, Cell Culture

Overexpression of human NRROS reduces the H 2 O 2 -induced ROS generation, IL-6 expression, and p47 phosphorylation. a RBA-1 cells were pretreated with 10 μM 15d-PGJ 2 for 1 h and then treated with 100 μM H 2 O 2 for the indicated time intervals. The levels of protein expression were analyzed by Western blotting. b RBA-1 cells were pretreated with 15d-PGJ 2 for 1 h and then treated with 100 μM H 2 O 2 for the indicated time intervals. The levels of IL-6 mRNA expression were determined by RT/real-time PCR. c , d RBA-1 stable clones of hNRROS and pCMV-Tag2B, or RBA-1 cells were treated with 100 μM H 2 O 2 for 1 h. c The levels of hNRROS mRNA expression were determined by RT/real-time PCR. d The cells were labeled with 10 μM H 2 DCFDA and then incubated with 100 μM H 2 O 2 for 30 min. The levels of ROS generation were observed using a fluorescence microscope. Scale bar = 100 μm. e RBA-1 stable clones of hNRROS and pCMV-Tag2B, or RBA-1 cells were treated with 100 μM H 2 O 2 for the indicated time intervals. The levels of protein expression were analyzed by Western blotting. f , g RBA-1 stable clones were treated with H 2 O 2 for the indicated time intervals. The levels of IL-6 mRNA expression and secretion of IL-6 were analyzed by RT/real-time PCR and ELISA kit, respectively. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). * p < 0.05, as compared with the respective values significantly different as indicated

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: Overexpression of human NRROS reduces the H 2 O 2 -induced ROS generation, IL-6 expression, and p47 phosphorylation. a RBA-1 cells were pretreated with 10 μM 15d-PGJ 2 for 1 h and then treated with 100 μM H 2 O 2 for the indicated time intervals. The levels of protein expression were analyzed by Western blotting. b RBA-1 cells were pretreated with 15d-PGJ 2 for 1 h and then treated with 100 μM H 2 O 2 for the indicated time intervals. The levels of IL-6 mRNA expression were determined by RT/real-time PCR. c , d RBA-1 stable clones of hNRROS and pCMV-Tag2B, or RBA-1 cells were treated with 100 μM H 2 O 2 for 1 h. c The levels of hNRROS mRNA expression were determined by RT/real-time PCR. d The cells were labeled with 10 μM H 2 DCFDA and then incubated with 100 μM H 2 O 2 for 30 min. The levels of ROS generation were observed using a fluorescence microscope. Scale bar = 100 μm. e RBA-1 stable clones of hNRROS and pCMV-Tag2B, or RBA-1 cells were treated with 100 μM H 2 O 2 for the indicated time intervals. The levels of protein expression were analyzed by Western blotting. f , g RBA-1 stable clones were treated with H 2 O 2 for the indicated time intervals. The levels of IL-6 mRNA expression and secretion of IL-6 were analyzed by RT/real-time PCR and ELISA kit, respectively. The data are presented as mean ± SEM, from three independent experiments ( n = 3, number of independent cell culture preparations). * p < 0.05, as compared with the respective values significantly different as indicated

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Over Expression, Expressing, Phospho-proteomics, Western Blot, Real-time Polymerase Chain Reaction, Clone Assay, Labeling, Incubation, Fluorescence, Microscopy, Enzyme-linked Immunosorbent Assay, Cell Culture

Scheme of signaling pathways is involved in 15d-PGJ 2 -induced NRROS expression suppressing the H 2 O 2 -induced IL-6 expression and ROS generation in RBA-1 cells. NRROS expression induced by 15d-PGJ 2 is mediated through a PPARγ-independent activation of PI3K/AKT leading to phosphorylation of FoxO1 and Sp1. These activated transcription factors bind to NRROS promoters and enhance NRROS transcription in RBA-1 cells. Up-regulation of NRROS not only downregulates the H 2 O 2 -mediated ROS generation but also inhibits the expression of pro-inflammatory cytokine IL-6. These results elucidate the mechanisms underlying 15d-PGJ 2 -induced NRROS expression which might be a potential strategy for management of brain inflammatory and neurodegenerative diseases

Journal: Neurotoxicity Research

Article Title: Involvement of FoxO1, Sp1, and Nrf2 in Upregulation of Negative Regulator of ROS by 15d-PGJ 2 Attenuates H 2 O 2 -Induced IL-6 Expression in Rat Brain Astrocytes

doi: 10.1007/s12640-020-00318-6

Figure Lengend Snippet: Scheme of signaling pathways is involved in 15d-PGJ 2 -induced NRROS expression suppressing the H 2 O 2 -induced IL-6 expression and ROS generation in RBA-1 cells. NRROS expression induced by 15d-PGJ 2 is mediated through a PPARγ-independent activation of PI3K/AKT leading to phosphorylation of FoxO1 and Sp1. These activated transcription factors bind to NRROS promoters and enhance NRROS transcription in RBA-1 cells. Up-regulation of NRROS not only downregulates the H 2 O 2 -mediated ROS generation but also inhibits the expression of pro-inflammatory cytokine IL-6. These results elucidate the mechanisms underlying 15d-PGJ 2 -induced NRROS expression which might be a potential strategy for management of brain inflammatory and neurodegenerative diseases

Article Snippet: 15d-PGJ 2 and 15d-PGJ 2 antibody (Cat# ADI-915-043-100) were from Enzo (New York, NY, USA).

Techniques: Protein-Protein interactions, Expressing, Activation Assay, Phospho-proteomics

Comparison of neurological deficits score.

Journal: BioMed Research International

Article Title: 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model

doi: 10.1155/2015/864509

Figure Lengend Snippet: Comparison of neurological deficits score.

Article Snippet: Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively.

Techniques: Comparison

Evaluation of microglia activation.

Journal: BioMed Research International

Article Title: 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model

doi: 10.1155/2015/864509

Figure Lengend Snippet: Evaluation of microglia activation.

Article Snippet: Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively.

Techniques: Activation Assay

Expression level of TNF- α (ng/L).

Journal: BioMed Research International

Article Title: 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model

doi: 10.1155/2015/864509

Figure Lengend Snippet: Expression level of TNF- α (ng/L).

Article Snippet: Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively.

Techniques: Expressing

Expression level of IL-1 β (ng/L).

Journal: BioMed Research International

Article Title: 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model

doi: 10.1155/2015/864509

Figure Lengend Snippet: Expression level of IL-1 β (ng/L).

Article Snippet: Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively.

Techniques: Expressing

Percentage of apoptotic cells.

Journal: BioMed Research International

Article Title: 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model

doi: 10.1155/2015/864509

Figure Lengend Snippet: Percentage of apoptotic cells.

Article Snippet: Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively.

Techniques:

Quantification of infarcted volume of brain lesion and cerebral edema volume.

Journal: BioMed Research International

Article Title: 15d-PGJ2 Reduced Microglia Activation and Alleviated Neurological Deficit of Ischemic Reperfusion in Diabetic Rat Model

doi: 10.1155/2015/864509

Figure Lengend Snippet: Quantification of infarcted volume of brain lesion and cerebral edema volume.

Article Snippet: Streptozotocin (STZ, Cat. number S0130) and 15d-PGJ2 (Cat. number 87893-55-8) were from Sigma (St. Louis, MO, United States) and Santa Cruz Biotechnology (Shanghai) Co., Ltd., respectively.

Techniques: