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Santa Cruz Biotechnology anti lamin a c
Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD.  A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis.  B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin.  C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P
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1) Product Images from "Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling"

Article Title: Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling

Journal: The FASEB Journal

doi: 10.1096/fj.201601155R

Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD.  A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis.  B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin.  C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P
Figure Legend Snippet: Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD. A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis. B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin. C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P

Techniques Used: Activation Assay, Western Blot, Translocation Assay, CTL Assay

Effect of FPS-ZM1 on CSE-induced activation of DAMP signaling in MLE-12 cells.  A – C ) After exposure to CSE in the presence or absence of FPS-ZM1 for 24 h, Western blot analysis was perfomed ( A ), and the expression levels of mRAGE in lung homogenates ( B ) and of circulating sRAGE ( C ) were determined by ELISA.  D ,  E ) IKKα, IKKβ, and the phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) ( D ) and the phosphorylation of JNK/ERK/p38 MAPK activation ( E ) were detected with the appropriate antibodies. β-Actin, total RelA/p65, total JNK1/2, total ERK1/2, and total p38 were used for loading controls.  F ) Correlation to Nrf2 translocation from cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for nuclear protein loading control, and β-actin was used for cytoplasmic protein loading control. The activation of DAMP signaling was inhibited by FPS-ZM1 treatment. Results are combined data from 3 independent experiments. Ctl, control group; ns, not significant. * P
Figure Legend Snippet: Effect of FPS-ZM1 on CSE-induced activation of DAMP signaling in MLE-12 cells. A – C ) After exposure to CSE in the presence or absence of FPS-ZM1 for 24 h, Western blot analysis was perfomed ( A ), and the expression levels of mRAGE in lung homogenates ( B ) and of circulating sRAGE ( C ) were determined by ELISA. D , E ) IKKα, IKKβ, and the phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) ( D ) and the phosphorylation of JNK/ERK/p38 MAPK activation ( E ) were detected with the appropriate antibodies. β-Actin, total RelA/p65, total JNK1/2, total ERK1/2, and total p38 were used for loading controls. F ) Correlation to Nrf2 translocation from cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for nuclear protein loading control, and β-actin was used for cytoplasmic protein loading control. The activation of DAMP signaling was inhibited by FPS-ZM1 treatment. Results are combined data from 3 independent experiments. Ctl, control group; ns, not significant. * P

Techniques Used: Activation Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Translocation Assay, CTL Assay

2) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Activation Assay, Transfection, Western Blot, Infection, Expressing, MTT Assay

3) Product Images from "Methylation of the KEAP1 gene promoter region in human colorectal cancer"

Article Title: Methylation of the KEAP1 gene promoter region in human colorectal cancer

Journal: BMC Cancer

doi: 10.1186/1471-2407-12-66

Expression of the Nrf2 target genes NQO- 1 and AKR1C1 after t-BHQ treatment . Real-time PCR analysis of the Nrf2 target genes NQO-1 ( A , left) and AKRC1 ( B , left) in HT29 cells (methylated) and Colo320DM cells (unmethylated). Cells were treated with the Keap1 stimulator t-BHQ for 24 h. Columns, mean ( n = 3); bars, SD. * P
Figure Legend Snippet: Expression of the Nrf2 target genes NQO- 1 and AKR1C1 after t-BHQ treatment . Real-time PCR analysis of the Nrf2 target genes NQO-1 ( A , left) and AKRC1 ( B , left) in HT29 cells (methylated) and Colo320DM cells (unmethylated). Cells were treated with the Keap1 stimulator t-BHQ for 24 h. Columns, mean ( n = 3); bars, SD. * P

Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Methylation

4) Product Images from "Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels"

Article Title: Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels

Journal: Scientific Reports

doi: 10.1038/s41598-018-26049-5

Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p
Figure Legend Snippet: Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p

Techniques Used: Expressing

5) Product Images from "Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity"

Article Title: Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0138771

Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P
Figure Legend Snippet: Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P

Techniques Used: Negative Control, Incubation, MTT Assay

6) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Activation Assay, Transfection, Western Blot, Infection, Expressing, MTT Assay

7) Product Images from "Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity"

Article Title: Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0138771

Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P
Figure Legend Snippet: Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P

Techniques Used: Negative Control, Incubation, MTT Assay

8) Product Images from "Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling"

Article Title: Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling

Journal: The FASEB Journal

doi: 10.1096/fj.201601155R

Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD. A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis. B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin. C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P
Figure Legend Snippet: Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD. A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis. B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin. C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P

Techniques Used: Activation Assay, Western Blot, Translocation Assay, CTL Assay

Effect of FPS-ZM1 on CSE-induced activation of DAMP signaling in MLE-12 cells. A – C ) After exposure to CSE in the presence or absence of FPS-ZM1 for 24 h, Western blot analysis was perfomed ( A ), and the expression levels of mRAGE in lung homogenates ( B ) and of circulating sRAGE ( C ) were determined by ELISA. D , E ) IKKα, IKKβ, and the phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) ( D ) and the phosphorylation of JNK/ERK/p38 MAPK activation ( E ) were detected with the appropriate antibodies. β-Actin, total RelA/p65, total JNK1/2, total ERK1/2, and total p38 were used for loading controls. F ) Correlation to Nrf2 translocation from cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for nuclear protein loading control, and β-actin was used for cytoplasmic protein loading control. The activation of DAMP signaling was inhibited by FPS-ZM1 treatment. Results are combined data from 3 independent experiments. Ctl, control group; ns, not significant. * P
Figure Legend Snippet: Effect of FPS-ZM1 on CSE-induced activation of DAMP signaling in MLE-12 cells. A – C ) After exposure to CSE in the presence or absence of FPS-ZM1 for 24 h, Western blot analysis was perfomed ( A ), and the expression levels of mRAGE in lung homogenates ( B ) and of circulating sRAGE ( C ) were determined by ELISA. D , E ) IKKα, IKKβ, and the phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) ( D ) and the phosphorylation of JNK/ERK/p38 MAPK activation ( E ) were detected with the appropriate antibodies. β-Actin, total RelA/p65, total JNK1/2, total ERK1/2, and total p38 were used for loading controls. F ) Correlation to Nrf2 translocation from cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for nuclear protein loading control, and β-actin was used for cytoplasmic protein loading control. The activation of DAMP signaling was inhibited by FPS-ZM1 treatment. Results are combined data from 3 independent experiments. Ctl, control group; ns, not significant. * P

Techniques Used: Activation Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Translocation Assay, CTL Assay

9) Product Images from "Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels"

Article Title: Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels

Journal: Scientific Reports

doi: 10.1038/s41598-018-26049-5

Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f ) Western blot analysis of protein bands. Uncropped immunoblot scans are presented in Supplementary Figure S1 . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p
Figure Legend Snippet: Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f ) Western blot analysis of protein bands. Uncropped immunoblot scans are presented in Supplementary Figure S1 . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p

Techniques Used: Expressing, Western Blot

Cytosine methylation analysis of chicken spermatozoa. Cytosine methylation of ( a ) NRF 2, ( b ) KEAP1 , ( c ) PRDX4 , ( d ) ATP5A1 , ( e ) AMPKα2 , and ( f ) mTOR were analyzed using CyMATE. Lengths of sequenced regions and position of cytosine are shown schematically. The order of the individual sequences of ten clones is listed on the left. 0 represents the control group; 11.7 represents plasma-treated group at 11.7 kV for 20 s; and 27.6 represents plasma-treated group at 27.6 kV for 20 s. The reference sequence is shown in the first line. The sequence is distinguished by circles for CG, squares for CHG, and triangles for CHH. Filled symbols represent methylated cytosine, and open symbols represent unmethylated cytosine. Average methylation levels for CG, CHG, and CHH of ( g ) NRF2 , ( h ) KEAP1 , ( i ) PRDX4 , ( j ) ATP5A1 , ( k ) AMPKα2 , and ( l ) mTOR .
Figure Legend Snippet: Cytosine methylation analysis of chicken spermatozoa. Cytosine methylation of ( a ) NRF 2, ( b ) KEAP1 , ( c ) PRDX4 , ( d ) ATP5A1 , ( e ) AMPKα2 , and ( f ) mTOR were analyzed using CyMATE. Lengths of sequenced regions and position of cytosine are shown schematically. The order of the individual sequences of ten clones is listed on the left. 0 represents the control group; 11.7 represents plasma-treated group at 11.7 kV for 20 s; and 27.6 represents plasma-treated group at 27.6 kV for 20 s. The reference sequence is shown in the first line. The sequence is distinguished by circles for CG, squares for CHG, and triangles for CHH. Filled symbols represent methylated cytosine, and open symbols represent unmethylated cytosine. Average methylation levels for CG, CHG, and CHH of ( g ) NRF2 , ( h ) KEAP1 , ( i ) PRDX4 , ( j ) ATP5A1 , ( k ) AMPKα2 , and ( l ) mTOR .

Techniques Used: Methylation, Clone Assay, Sequencing

10) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.
Figure Legend Snippet: Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.

Techniques Used: Transfection, Western Blot

PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Activation Assay, Transfection, Western Blot, Infection, Expressing, MTT Assay

11) Product Images from "The Cinnamon-derived Dietary Factor Cinnamic Aldehyde Activates the Nrf2-dependent Antioxidant Response in Human Epithelial Colon Cells"

Article Title: The Cinnamon-derived Dietary Factor Cinnamic Aldehyde Activates the Nrf2-dependent Antioxidant Response in Human Epithelial Colon Cells

Journal: Molecules (Basel, Switzerland)

doi: 10.3390/molecules15053338

Upregulation of Nrf2 and HO-1 protein levels in normal human fetal epithelial colon cells exposed to cinnamic aldehyde and ethanolic cinnamon extract. FHC cells were treated with either CA (10 μM), CE (dose CA: 10 μM), or tBHQ (100 μM) for 4 h. Equal loading was assessed by immunodetection of lamin A.
Figure Legend Snippet: Upregulation of Nrf2 and HO-1 protein levels in normal human fetal epithelial colon cells exposed to cinnamic aldehyde and ethanolic cinnamon extract. FHC cells were treated with either CA (10 μM), CE (dose CA: 10 μM), or tBHQ (100 μM) for 4 h. Equal loading was assessed by immunodetection of lamin A.

Techniques Used: Immunodetection

Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and Keap1 proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p
Figure Legend Snippet: Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and Keap1 proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p

Techniques Used: Activation Assay, Multiple Displacement Amplification, Luciferase, Expressing, Plasmid Preparation, Transfection, Activity Assay

Upregulation of Nrf2 protein levels in cultured human colon cancer cells exposed to cinnamic aldehyde and ethanolic cinnamon extract. Cultured human HCT116 ( a ) and HT29 cells ( b ) were treated for 4 h with either CA or CE at the indicated doses normalized for CA content. Treatment with tBHQ (100 μM, 4h) served as a positive control. Equal loading was assessed by immunodetection of β-actin.
Figure Legend Snippet: Upregulation of Nrf2 protein levels in cultured human colon cancer cells exposed to cinnamic aldehyde and ethanolic cinnamon extract. Cultured human HCT116 ( a ) and HT29 cells ( b ) were treated for 4 h with either CA or CE at the indicated doses normalized for CA content. Treatment with tBHQ (100 μM, 4h) served as a positive control. Equal loading was assessed by immunodetection of β-actin.

Techniques Used: Cell Culture, Positive Control, Immunodetection

12) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

PI activates IκB/NF-κB pathway (A) NCI-H157 and A549 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for IκBα and GAPDH. (B) NCI-H157 cells were treated with PS-341 or MG132 for the indicated times. Nuclear proteins were extracted and subjected to Western blot analysis for p65 and Lamin A/C. TNF-α (10 ng/mL, 60 min) was used as a positive control. (C) Both cell lines were stimulated with PS-341 or MG132 for 0, 6, 8, 12, or 24 h. Total RNA was isolated and quantitative real-time PCR for COX-2 and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI activates IκB/NF-κB pathway (A) NCI-H157 and A549 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for IκBα and GAPDH. (B) NCI-H157 cells were treated with PS-341 or MG132 for the indicated times. Nuclear proteins were extracted and subjected to Western blot analysis for p65 and Lamin A/C. TNF-α (10 ng/mL, 60 min) was used as a positive control. (C) Both cell lines were stimulated with PS-341 or MG132 for 0, 6, 8, 12, or 24 h. Total RNA was isolated and quantitative real-time PCR for COX-2 and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Western Blot, Positive Control, Isolation, Real-time Polymerase Chain Reaction

13) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.
Figure Legend Snippet: Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.

Techniques Used: Transfection, Western Blot

14) Product Images from "A plastic SQSTM1/p62-dependent autophagic reserve maintains proteostasis and determines proteasome inhibitor susceptibility in multiple myeloma cells"

Article Title: A plastic SQSTM1/p62-dependent autophagic reserve maintains proteostasis and determines proteasome inhibitor susceptibility in multiple myeloma cells

Journal: Autophagy

doi: 10.1080/15548627.2015.1052928

Acute proteasome stress co-opts SQSTM1 onto protein aggregates, and induces de novo SQSTM1 expression. ( A, B ) Immunofluorescence analysis of SQSTM1 and ubiquitin accumulation in MM lines upon treatment with bortezomib (Btz). Nuclei are stained blue with DAPI. Scale bars: 10 µm. ( A ) SQSTM1 in MM.1S cells left untreated (left) or treated for 1 h with 1 µM Btz (right) (n = 5 independent experiments). ( B ) SQSTM1 and ubiquitin in MM.1S cells treated with Btz (as in A ). ( C ) Co-immunoprecipitation (IP) of polyubiquitinated proteins with SQSTM1. MM.1S cells were treated with Btz (as in A ), prior to IP of SQSTM1, and the association of ubiquitinated proteins and KEAP1 with SQSTM1 assessed by immunoblot (n ≥ 3 ). ( D ) Changes of selected proteins of the SQSTM1 interactome upon treatment with Btz (as in A ) as determined by SILAC LC-MS/MS in MM.1S cells (more proteins listed in Table 1 ). ( E ) Quantitative RT-PCR analysis of transcripts encoding the indicated autophagy receptors in MM lines treated with 1 µM Btz for 4 h. mRNA amounts were normalized by histone H3 and expressed relative to untreated controls (average induction ±s.e.m.; n = 3). ( F ) Immunoblot analysis of SQSTM1 and LC3 in the indicated MM lines treated with 1 µM Btz for 8 h (representative blot, n = 3). ( G ) Immunoblot analysis of SQSTM1 in MM.1S cells treated with 1 µM Btz for 8 h in the presence or absence of 10 µg/ml cycloheximide (CHX).
Figure Legend Snippet: Acute proteasome stress co-opts SQSTM1 onto protein aggregates, and induces de novo SQSTM1 expression. ( A, B ) Immunofluorescence analysis of SQSTM1 and ubiquitin accumulation in MM lines upon treatment with bortezomib (Btz). Nuclei are stained blue with DAPI. Scale bars: 10 µm. ( A ) SQSTM1 in MM.1S cells left untreated (left) or treated for 1 h with 1 µM Btz (right) (n = 5 independent experiments). ( B ) SQSTM1 and ubiquitin in MM.1S cells treated with Btz (as in A ). ( C ) Co-immunoprecipitation (IP) of polyubiquitinated proteins with SQSTM1. MM.1S cells were treated with Btz (as in A ), prior to IP of SQSTM1, and the association of ubiquitinated proteins and KEAP1 with SQSTM1 assessed by immunoblot (n ≥ 3 ). ( D ) Changes of selected proteins of the SQSTM1 interactome upon treatment with Btz (as in A ) as determined by SILAC LC-MS/MS in MM.1S cells (more proteins listed in Table 1 ). ( E ) Quantitative RT-PCR analysis of transcripts encoding the indicated autophagy receptors in MM lines treated with 1 µM Btz for 4 h. mRNA amounts were normalized by histone H3 and expressed relative to untreated controls (average induction ±s.e.m.; n = 3). ( F ) Immunoblot analysis of SQSTM1 and LC3 in the indicated MM lines treated with 1 µM Btz for 8 h (representative blot, n = 3). ( G ) Immunoblot analysis of SQSTM1 in MM.1S cells treated with 1 µM Btz for 8 h in the presence or absence of 10 µg/ml cycloheximide (CHX).

Techniques Used: Expressing, Immunofluorescence, Staining, Immunoprecipitation, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Quantitative RT-PCR

15) Product Images from "Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels"

Article Title: Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels

Journal: Scientific Reports

doi: 10.1038/s41598-018-26049-5

Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p
Figure Legend Snippet: Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p

Techniques Used: Expressing

16) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.
Figure Legend Snippet: Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.

Techniques Used: Transfection, Western Blot

17) Product Images from "Schisandrol B protects against acetaminophen-induced acute hepatotoxicity in mice via activation of the NRF2/ARE signaling pathway"

Article Title: Schisandrol B protects against acetaminophen-induced acute hepatotoxicity in mice via activation of the NRF2/ARE signaling pathway

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2015.120

The effects of SolB on activation and expression of NRF2 in vivo . (A) qRT-PCR analysis was performed to measure the gene expression of Nrf2, and the data are expressed as the mean±SEM ( n =6). (B–C) Western blotting was used to measure nuclear NRF2 protein expression. Specific band intensities were quantified and normalized to histone H3 and expressed as the mean±SEM ( n =3). (D–E) Western blotting was used to measure cytoplasmic KEAP1 protein expression. Specific band intensities were quantified and normalized to GAPDH and expressed as the mean±SEM ( n =3). b P
Figure Legend Snippet: The effects of SolB on activation and expression of NRF2 in vivo . (A) qRT-PCR analysis was performed to measure the gene expression of Nrf2, and the data are expressed as the mean±SEM ( n =6). (B–C) Western blotting was used to measure nuclear NRF2 protein expression. Specific band intensities were quantified and normalized to histone H3 and expressed as the mean±SEM ( n =3). (D–E) Western blotting was used to measure cytoplasmic KEAP1 protein expression. Specific band intensities were quantified and normalized to GAPDH and expressed as the mean±SEM ( n =3). b P

Techniques Used: Activation Assay, Expressing, In Vivo, Quantitative RT-PCR, Western Blot

Role of NRF2 signaling and its downstream proteins in SolB-mediated protection against APAP-induced liver toxicity. (A) Western blotting was used to measure the protein expression of GSR, GSS, GCLC, GCLM, GST-α, GST-μ, GST-π, NQO1, HO-1, MRP2, MRP3 and MRP4. (B–M) Specific band intensities were quantified and normalized to GAPDH. The data are expressed as the mean±SEM ( n =3). b P
Figure Legend Snippet: Role of NRF2 signaling and its downstream proteins in SolB-mediated protection against APAP-induced liver toxicity. (A) Western blotting was used to measure the protein expression of GSR, GSS, GCLC, GCLM, GST-α, GST-μ, GST-π, NQO1, HO-1, MRP2, MRP3 and MRP4. (B–M) Specific band intensities were quantified and normalized to GAPDH. The data are expressed as the mean±SEM ( n =3). b P

Techniques Used: Western Blot, Expressing

Effects of SolB on cell viability and NRF2 activation. (A) An MTT assay was used to measure HepG2 cell viability after treatment with SolB at 2.5 to 160 μmol/L. (B) A luciferase reporter assay was used to measure the effect of SolB on NRF2 activation. The HepG2 cells were transiently transfected with plasmids as described in the Materials and methods section, and 6 h later the cells were treated with various concentrations of SolB (2.5-20 μmol/L) or the positive agonist SFN (10 μmol/L) for 24 h. The data are expressed as the mean±SEM ( n =5). c P
Figure Legend Snippet: Effects of SolB on cell viability and NRF2 activation. (A) An MTT assay was used to measure HepG2 cell viability after treatment with SolB at 2.5 to 160 μmol/L. (B) A luciferase reporter assay was used to measure the effect of SolB on NRF2 activation. The HepG2 cells were transiently transfected with plasmids as described in the Materials and methods section, and 6 h later the cells were treated with various concentrations of SolB (2.5-20 μmol/L) or the positive agonist SFN (10 μmol/L) for 24 h. The data are expressed as the mean±SEM ( n =5). c P

Techniques Used: Activation Assay, MTT Assay, Luciferase, Reporter Assay, Transfection

18) Product Images from "Protective Effects of Berberine on Renal Injury in Streptozotocin (STZ)-Induced Diabetic Mice"

Article Title: Protective Effects of Berberine on Renal Injury in Streptozotocin (STZ)-Induced Diabetic Mice

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms17081327

BBR enhanced activation of Nrf2/HO-1 signaling in diabetic kidneys. Representative western blots demonstrated that BBR treatment increased protein expression of Nrf2, NQO1 and HO-1 in the kidneys of diabetic mice ( n = 8). Each value represents mean ± SEM. All experiments were performed three times with virtually identical results. β-actin was used as loading control. (** p
Figure Legend Snippet: BBR enhanced activation of Nrf2/HO-1 signaling in diabetic kidneys. Representative western blots demonstrated that BBR treatment increased protein expression of Nrf2, NQO1 and HO-1 in the kidneys of diabetic mice ( n = 8). Each value represents mean ± SEM. All experiments were performed three times with virtually identical results. β-actin was used as loading control. (** p

Techniques Used: Activation Assay, Western Blot, Expressing, Mouse Assay

Antifibrotic effects of BBR were associated with the inhibition of EMT in diabetic kidneys. Representative western blots of α-SMA and collagen-I in the kidneys of control mice, diabetic mice or BBR-treated diabetic mice. All experiments were performed three times with virtually identical results. β-actin was used as loading control. (** p
Figure Legend Snippet: Antifibrotic effects of BBR were associated with the inhibition of EMT in diabetic kidneys. Representative western blots of α-SMA and collagen-I in the kidneys of control mice, diabetic mice or BBR-treated diabetic mice. All experiments were performed three times with virtually identical results. β-actin was used as loading control. (** p

Techniques Used: Inhibition, Western Blot, Mouse Assay

Influence of BBR on HG-induced Nrf2/HO-1 signaling in NRK-52E cells . The NRK-52E cells were treated with 30 mM HG for 48 h, in the presence or absence of BBR. The expression NQO1, HO-1 and Nrf2 was analyzed using western blot ( n = 6). ( a – e ) The results were representatives of three independent experiments. β-actin was used as loading control. (** p
Figure Legend Snippet: Influence of BBR on HG-induced Nrf2/HO-1 signaling in NRK-52E cells . The NRK-52E cells were treated with 30 mM HG for 48 h, in the presence or absence of BBR. The expression NQO1, HO-1 and Nrf2 was analyzed using western blot ( n = 6). ( a – e ) The results were representatives of three independent experiments. β-actin was used as loading control. (** p

Techniques Used: Expressing, Western Blot

19) Product Images from "Effect of p62 on tau hyperphosphorylation in a rat model of Alzheimer's disease ☆"

Article Title: Effect of p62 on tau hyperphosphorylation in a rat model of Alzheimer's disease ☆

Journal: Neural Regeneration Research

doi: 10.3969/j.issn.1673-5374.2012.17.004

Changes in Keap1 and Nrf2 expression in the rat cerebral cortex and hippocampus. (A, B) Immunohistochemical and quantitative analysis for the expression of Keap1 in the cerebral cortex of Alzheimer's disease (AD) model rats. The expression of Keap1 in AD model rats was increased compared with the control (CON) group. (C, D) Immunohistochemical and quantitative analysis for the expression of Nrf2 in the cerebral cortex of AD model rats. The expression of Nrf2 in AD model rats increased compared with control rats. (E, F) Immunohistochemical and quantitative analysis for the expression of Keap1 in the hippocampus of AD model rats. The expression of Keap1 in AD model rats increased compared with control rats. (G, H) Immunohistochemical and quantitative analysis for the expression of Nrf2 in the hippocampus of AD model rats. The expression of Nrf2 in AD model rats remained unchanged compared with control rats. Scale bars: 20 μm. a P
Figure Legend Snippet: Changes in Keap1 and Nrf2 expression in the rat cerebral cortex and hippocampus. (A, B) Immunohistochemical and quantitative analysis for the expression of Keap1 in the cerebral cortex of Alzheimer's disease (AD) model rats. The expression of Keap1 in AD model rats was increased compared with the control (CON) group. (C, D) Immunohistochemical and quantitative analysis for the expression of Nrf2 in the cerebral cortex of AD model rats. The expression of Nrf2 in AD model rats increased compared with control rats. (E, F) Immunohistochemical and quantitative analysis for the expression of Keap1 in the hippocampus of AD model rats. The expression of Keap1 in AD model rats increased compared with control rats. (G, H) Immunohistochemical and quantitative analysis for the expression of Nrf2 in the hippocampus of AD model rats. The expression of Nrf2 in AD model rats remained unchanged compared with control rats. Scale bars: 20 μm. a P

Techniques Used: Expressing, Immunohistochemistry

20) Product Images from "Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells"

Article Title: Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2015.2725

Graphical representation of mRNA quantification of antioxidants Nrf2 and Keap1 in hLECs cultured for 24 h with 0, 1, 4 and 20% oxygen. *P
Figure Legend Snippet: Graphical representation of mRNA quantification of antioxidants Nrf2 and Keap1 in hLECs cultured for 24 h with 0, 1, 4 and 20% oxygen. *P

Techniques Used: Cell Culture

21) Product Images from "MiR-28 regulates Nrf2 expression through a Keap1-independent mechanism"

Article Title: MiR-28 regulates Nrf2 expression through a Keap1-independent mechanism

Journal: Breast cancer research and treatment

doi: 10.1007/s10549-011-1604-1

miR-28 regulation of Nrf2 expression is Keap1 independent. a MCF-7 cells were co-transfected with Keap1-FLAG and pri-miR-28 or vehicle-control. Western blotting was used to detect Keap1 expression with anti-FLAG antibody. β -actin was used as a loading control. b Keap1-FLAG and Nrf2-myc were co-transfected into HEK293T cells with pri-miR-28 or vehicle control. Co-immunoprecipitation was performed to examine Nrf2-Keap1 interaction status. The anti-myc antibody was used to pull down antigens and the mouse IgG was used as negative control; anti-FLAG or anti-myc antibody was used for Western blotting. A representative experiment was shown. Normalized protein expression in input was analyzed using UN-SCAN-IT program
Figure Legend Snippet: miR-28 regulation of Nrf2 expression is Keap1 independent. a MCF-7 cells were co-transfected with Keap1-FLAG and pri-miR-28 or vehicle-control. Western blotting was used to detect Keap1 expression with anti-FLAG antibody. β -actin was used as a loading control. b Keap1-FLAG and Nrf2-myc were co-transfected into HEK293T cells with pri-miR-28 or vehicle control. Co-immunoprecipitation was performed to examine Nrf2-Keap1 interaction status. The anti-myc antibody was used to pull down antigens and the mouse IgG was used as negative control; anti-FLAG or anti-myc antibody was used for Western blotting. A representative experiment was shown. Normalized protein expression in input was analyzed using UN-SCAN-IT program

Techniques Used: Expressing, Transfection, Western Blot, Immunoprecipitation, Negative Control

22) Product Images from "Protective Effects of Berberine on Renal Injury in Streptozotocin (STZ)-Induced Diabetic Mice"

Article Title: Protective Effects of Berberine on Renal Injury in Streptozotocin (STZ)-Induced Diabetic Mice

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms17081327

siRNA knockdown of Nrf2 abrogates BBR-induced NQO1 and HO-1 expression. ( a ) NRK-52E cells were transfected with Nrf2-siRNA and western blot analysis was performed with an antibody against Nrf2 was performed at various time-points following transfection (24, 48 and 72 h). Relative Nrf2 expression levels were calculated and normalized to the loading control. Corresponding protein levels were assessed using densitometry and expressed in relative intensities. All results were obtained from three independent experiments. Values are expressed as the mean ± SEM ( n = 6; ** p
Figure Legend Snippet: siRNA knockdown of Nrf2 abrogates BBR-induced NQO1 and HO-1 expression. ( a ) NRK-52E cells were transfected with Nrf2-siRNA and western blot analysis was performed with an antibody against Nrf2 was performed at various time-points following transfection (24, 48 and 72 h). Relative Nrf2 expression levels were calculated and normalized to the loading control. Corresponding protein levels were assessed using densitometry and expressed in relative intensities. All results were obtained from three independent experiments. Values are expressed as the mean ± SEM ( n = 6; ** p

Techniques Used: Expressing, Transfection, Western Blot

BBR enhanced activation of Nrf2/HO-1 signaling in diabetic kidneys. Representative western blots demonstrated that BBR treatment increased protein expression of Nrf2, NQO1 and HO-1 in the kidneys of diabetic mice ( n = 8). Each value represents mean ± SEM. All experiments were performed three times with virtually identical results. β-actin was used as loading control. (** p
Figure Legend Snippet: BBR enhanced activation of Nrf2/HO-1 signaling in diabetic kidneys. Representative western blots demonstrated that BBR treatment increased protein expression of Nrf2, NQO1 and HO-1 in the kidneys of diabetic mice ( n = 8). Each value represents mean ± SEM. All experiments were performed three times with virtually identical results. β-actin was used as loading control. (** p

Techniques Used: Activation Assay, Western Blot, Expressing, Mouse Assay

Influence of BBR on HG-induced Nrf2/HO-1 signaling in NRK-52E cells . The NRK-52E cells were treated with 30 mM HG for 48 h, in the presence or absence of BBR. The expression NQO1, HO-1 and Nrf2 was analyzed using western blot ( n = 6). ( a – e ) The results were representatives of three independent experiments. β-actin was used as loading control. (** p
Figure Legend Snippet: Influence of BBR on HG-induced Nrf2/HO-1 signaling in NRK-52E cells . The NRK-52E cells were treated with 30 mM HG for 48 h, in the presence or absence of BBR. The expression NQO1, HO-1 and Nrf2 was analyzed using western blot ( n = 6). ( a – e ) The results were representatives of three independent experiments. β-actin was used as loading control. (** p

Techniques Used: Expressing, Western Blot

Nrf2 is involved in the inhibitory effect of BBR on the TGF-β/Smad signaling pathway. NRK-52E cells were divided into six groups as mentioned above, corresponding protein levels of Smad2/3, phospho-Smad2/3 and EMT markers were assessed using densitometry and expressed in relative intensities. All results were obtained from three independent experiments. All results were obtained from three independent experiments. Values are expressed as the mean ± SEM ( n = 6) (* p
Figure Legend Snippet: Nrf2 is involved in the inhibitory effect of BBR on the TGF-β/Smad signaling pathway. NRK-52E cells were divided into six groups as mentioned above, corresponding protein levels of Smad2/3, phospho-Smad2/3 and EMT markers were assessed using densitometry and expressed in relative intensities. All results were obtained from three independent experiments. All results were obtained from three independent experiments. Values are expressed as the mean ± SEM ( n = 6) (* p

Techniques Used:

23) Product Images from "Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells"

Article Title: Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2015.2725

Western blot analysis of ROS- and apoptosis-related UPR proteins. (A) ROS-related and (B) apoptosis-related UPR proteins in the hLECs cultured for 24 h with 0, 1, 4 and 20% oxygen. ROS, reactive oxygen species; UPR, unfolded protein response; Ero1-L, endoplasmic reticulum oxidoreductin 1-like; PDI, protein disulfide isomerase; ATF4, activating transcription factor 4; CHOP, C/EBP homologous protein.
Figure Legend Snippet: Western blot analysis of ROS- and apoptosis-related UPR proteins. (A) ROS-related and (B) apoptosis-related UPR proteins in the hLECs cultured for 24 h with 0, 1, 4 and 20% oxygen. ROS, reactive oxygen species; UPR, unfolded protein response; Ero1-L, endoplasmic reticulum oxidoreductin 1-like; PDI, protein disulfide isomerase; ATF4, activating transcription factor 4; CHOP, C/EBP homologous protein.

Techniques Used: Western Blot, Cell Culture

24) Product Images from "Decreased DJ-1 Leads to Impaired Nrf2-Regulated Antioxidant Defense and Increased UV-A–Induced Apoptosis in Corneal Endothelial Cells"

Article Title: Decreased DJ-1 Leads to Impaired Nrf2-Regulated Antioxidant Defense and Increased UV-A–Induced Apoptosis in Corneal Endothelial Cells

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.14-14580

The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. ( A ) Left panel
Figure Legend Snippet: The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. ( A ) Left panel

Techniques Used: Immunoprecipitation, Western Blot

DJ-1 downregulation decreases Nrf2 nuclear translocation and its target gene expression. Nrf2 subcellular localization. ( A ) Representative Western blot of Nrf2 protein levels in the cytosolic and nuclear fractions of non–siRNA-treated, control
Figure Legend Snippet: DJ-1 downregulation decreases Nrf2 nuclear translocation and its target gene expression. Nrf2 subcellular localization. ( A ) Representative Western blot of Nrf2 protein levels in the cytosolic and nuclear fractions of non–siRNA-treated, control

Techniques Used: Translocation Assay, Expressing, Western Blot

25) Product Images from "Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity"

Article Title: Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0138771

Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P
Figure Legend Snippet: Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P

Techniques Used: Negative Control, Incubation, MTT Assay

26) Product Images from "The Cinnamon-derived Dietary Factor Cinnamic Aldehyde Activates the Nrf2-dependent Antioxidant Response in Human Epithelial Colon Cells"

Article Title: The Cinnamon-derived Dietary Factor Cinnamic Aldehyde Activates the Nrf2-dependent Antioxidant Response in Human Epithelial Colon Cells

Journal: Molecules (Basel, Switzerland)

doi: 10.3390/molecules15053338

Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and Keap1 proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p
Figure Legend Snippet: Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and Keap1 proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p

Techniques Used: Activation Assay, Multiple Displacement Amplification, Luciferase, Expressing, Plasmid Preparation, Transfection, Activity Assay

27) Product Images from "Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling"

Article Title: Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling

Journal: The FASEB Journal

doi: 10.1096/fj.201601155R

Proposed mechanism for the effect of FPS-ZM1 in COPD. Exposure to CSE and release of elastase by injured cells during smoking increase ligands, such as S100 calgranulins and HMGB1. The ligands bind to and activate response by RAGE. The engagement of RAGE is linked to the DAMP signaling pathway, notably NF-κB and MAPKs, which overlap considerably with the Nrf2 signaling pathway for RAGE-mediated emphysematous response to stress stimuli in the lung (airspace enlargement, sustained inflammation, excessive ROS/RNS, immune cell infiltration, and epithelial cell death). When mice exposed to PPE or epithelial cells exposed to CSE are treated with FPS-ZM1, RAGE-mediated emphysema and DAMP signaling pathway recover. However, FPS-ZM1 treatment of Nrf2-deficient mice after elastase application fails to repair airspace enlargement, excessive ROS/RNS, and activation of DAMP signaling, including NF-κB and MAPK signaling; FPS-ZM1 reverses only the infiltration of inflammatory cells and cytokines. Eventually, boosted RAGE expression promotes COPD progression. In this respect, FPS-ZM1 may offer a potential therapeutic application to inhibit inflammation during the development of COPD, where this sustained inflammation is a big hurdle. Taken together, there is a need to verify the affirmative action of FPS-ZM1 against excessive infiltration of inflammatory cells and cytokines in COPD.
Figure Legend Snippet: Proposed mechanism for the effect of FPS-ZM1 in COPD. Exposure to CSE and release of elastase by injured cells during smoking increase ligands, such as S100 calgranulins and HMGB1. The ligands bind to and activate response by RAGE. The engagement of RAGE is linked to the DAMP signaling pathway, notably NF-κB and MAPKs, which overlap considerably with the Nrf2 signaling pathway for RAGE-mediated emphysematous response to stress stimuli in the lung (airspace enlargement, sustained inflammation, excessive ROS/RNS, immune cell infiltration, and epithelial cell death). When mice exposed to PPE or epithelial cells exposed to CSE are treated with FPS-ZM1, RAGE-mediated emphysema and DAMP signaling pathway recover. However, FPS-ZM1 treatment of Nrf2-deficient mice after elastase application fails to repair airspace enlargement, excessive ROS/RNS, and activation of DAMP signaling, including NF-κB and MAPK signaling; FPS-ZM1 reverses only the infiltration of inflammatory cells and cytokines. Eventually, boosted RAGE expression promotes COPD progression. In this respect, FPS-ZM1 may offer a potential therapeutic application to inhibit inflammation during the development of COPD, where this sustained inflammation is a big hurdle. Taken together, there is a need to verify the affirmative action of FPS-ZM1 against excessive infiltration of inflammatory cells and cytokines in COPD.

Techniques Used: Mouse Assay, Activation Assay, Expressing

28) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.
Figure Legend Snippet: Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.

Techniques Used: Transfection, Western Blot

29) Product Images from "Decreased DJ-1 Leads to Impaired Nrf2-Regulated Antioxidant Defense and Increased UV-A–Induced Apoptosis in Corneal Endothelial Cells"

Article Title: Decreased DJ-1 Leads to Impaired Nrf2-Regulated Antioxidant Defense and Increased UV-A–Induced Apoptosis in Corneal Endothelial Cells

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.14-14580

The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. ( A ) Left panel
Figure Legend Snippet: The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. ( A ) Left panel

Techniques Used: Immunoprecipitation, Western Blot

30) Product Images from "Decreased DJ-1 Leads to Impaired Nrf2-Regulated Antioxidant Defense and Increased UV-A–Induced Apoptosis in Corneal Endothelial Cells"

Article Title: Decreased DJ-1 Leads to Impaired Nrf2-Regulated Antioxidant Defense and Increased UV-A–Induced Apoptosis in Corneal Endothelial Cells

Journal: Investigative Ophthalmology & Visual Science

doi: 10.1167/iovs.14-14580

The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. ( A ) Left panel
Figure Legend Snippet: The effect of DJ-1 downregulation on Nrf2-associated Cul3 and Keap1 levels. Immunoprecipitation–WB analysis of Cul3/Nrf2 and Keap/Nrf2 levels in non–siRNA-treated, control siRNA-treated, and DJ-1 siRNA-treated HCECi cells. ( A ) Left panel

Techniques Used: Immunoprecipitation, Western Blot

31) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

PI-induced IκBα degradation is associated with macroautophagy (A) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for Lamp2a, Lamp2, LC3B, and GAPDH. (B) Cells were stimulated with PS-341 (50 nM) for the indicated times. Total RNA was isolated and quantitative real-time PCR for LC3B and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced IκBα degradation is associated with macroautophagy (A) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for Lamp2a, Lamp2, LC3B, and GAPDH. (B) Cells were stimulated with PS-341 (50 nM) for the indicated times. Total RNA was isolated and quantitative real-time PCR for LC3B and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Western Blot, Isolation, Real-time Polymerase Chain Reaction

Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.
Figure Legend Snippet: Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.

Techniques Used: Transfection, Western Blot

32) Product Images from "Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells"

Article Title: Attenuation of oxygen fluctuation-induced endoplasmic reticulum stress in human lens epithelial cells

Journal: Experimental and Therapeutic Medicine

doi: 10.3892/etm.2015.2725

Western blot analysis of the protein levels of HIF)-1α hypoxia-inducible factor in hLECs cultured for 1, 6, 12 and 24 h with 0, 1, 4 and 20% oxygen. HIF, hypoxia-inducible factor; hLECs, human lens epithelial cells.
Figure Legend Snippet: Western blot analysis of the protein levels of HIF)-1α hypoxia-inducible factor in hLECs cultured for 1, 6, 12 and 24 h with 0, 1, 4 and 20% oxygen. HIF, hypoxia-inducible factor; hLECs, human lens epithelial cells.

Techniques Used: Western Blot, Cell Culture

33) Product Images from "Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels"

Article Title: Non-thermal plasma treatment improves chicken sperm motility via the regulation of demethylation levels

Journal: Scientific Reports

doi: 10.1038/s41598-018-26049-5

Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f ) Western blot analysis of protein bands. Uncropped immunoblot scans are presented in Supplementary Figure S1 . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p
Figure Legend Snippet: Effect of plasma on chicken sperm mRNA and protein expression. Semen of 60-week-old cocks was exposed to varying plasma potentials for 20 s. Relative mRNA levels of the following genes were measured: ( a ) NOX4 , NRF2 , and KEAP1 ; ( b ) SOD , CAT , and GPx ; ( c ) PRDX1 , PRDX 3, PRDX4 , and PRDX6 ; ( d ) ATP5A1 , ATP5B , ATP5C1 , ATP5F1 , ATP5G1 , ATP5G3 , ATP5H , ATP5I , ATP5J , ATP5J 2, ATP5L , and ATP5S ; and ( e ) AMPKα2 , AMPKβ2 , AMPKγ3 , and mTOR . ( f ) Western blot analysis of protein bands. Uncropped immunoblot scans are presented in Supplementary Figure S1 . The grouping of gels/blots cropped from different gels. All blots were visualized with 5 min exposure time. Relative protein levels of ( g ) NRF2, KEAP1, PRDX4, ( h ) ATP5A, ( i ) p-AMPKα/AMPKα, and ( j ) p-mTOR/mTOR. Values are expressed as the mean ± standard error (n = 3) of three replicates; n represents an individual cock. * p

Techniques Used: Expressing, Western Blot

Cytosine methylation analysis of chicken spermatozoa. Cytosine methylation of ( a ) NRF 2, ( b ) KEAP1 , ( c ) PRDX4 , ( d ) ATP5A1 , ( e ) AMPKα2 , and ( f ) mTOR were analyzed using CyMATE. Lengths of sequenced regions and position of cytosine are shown schematically. The order of the individual sequences of ten clones is listed on the left. 0 represents the control group; 11.7 represents plasma-treated group at 11.7 kV for 20 s; and 27.6 represents plasma-treated group at 27.6 kV for 20 s. The reference sequence is shown in the first line. The sequence is distinguished by circles for CG, squares for CHG, and triangles for CHH. Filled symbols represent methylated cytosine, and open symbols represent unmethylated cytosine. Average methylation levels for CG, CHG, and CHH of ( g ) NRF2 , ( h ) KEAP1 , ( i ) PRDX4 , ( j ) ATP5A1 , ( k ) AMPKα2 , and ( l ) mTOR .
Figure Legend Snippet: Cytosine methylation analysis of chicken spermatozoa. Cytosine methylation of ( a ) NRF 2, ( b ) KEAP1 , ( c ) PRDX4 , ( d ) ATP5A1 , ( e ) AMPKα2 , and ( f ) mTOR were analyzed using CyMATE. Lengths of sequenced regions and position of cytosine are shown schematically. The order of the individual sequences of ten clones is listed on the left. 0 represents the control group; 11.7 represents plasma-treated group at 11.7 kV for 20 s; and 27.6 represents plasma-treated group at 27.6 kV for 20 s. The reference sequence is shown in the first line. The sequence is distinguished by circles for CG, squares for CHG, and triangles for CHH. Filled symbols represent methylated cytosine, and open symbols represent unmethylated cytosine. Average methylation levels for CG, CHG, and CHH of ( g ) NRF2 , ( h ) KEAP1 , ( i ) PRDX4 , ( j ) ATP5A1 , ( k ) AMPKα2 , and ( l ) mTOR .

Techniques Used: Methylation, Clone Assay, Sequencing

34) Product Images from "Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells"

Article Title: Proteasome Inhibitor-Induced IκB/NF-κB Activation is Mediated by Nrf2-Dependent Light Chain 3B Induction in Lung Cancer Cells

Journal: Molecules and Cells

doi: 10.14348/molcells.2018.0277

PI-induced IκBα degradation is mediated by Nrf2 up-regulation via both de novo protein synthesis and KEAP1 degradation (A, B) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for phospho-mTOR (Ser2448) (p-mTOR), sirtuin1, Nrf2, KEAP1, and GAPDH (A). Total RNA was isolated and quantitative real-time PCR for Nrf2 and GAPDH was performed (B). Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced IκBα degradation is mediated by Nrf2 up-regulation via both de novo protein synthesis and KEAP1 degradation (A, B) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for phospho-mTOR (Ser2448) (p-mTOR), sirtuin1, Nrf2, KEAP1, and GAPDH (A). Total RNA was isolated and quantitative real-time PCR for Nrf2 and GAPDH was performed (B). Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Western Blot, Isolation, Real-time Polymerase Chain Reaction

PI-induced IκBα degradation is associated with macroautophagy (A) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for Lamp2a, Lamp2, LC3B, and GAPDH. (B) Cells were stimulated with PS-341 (50 nM) for the indicated times. Total RNA was isolated and quantitative real-time PCR for LC3B and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced IκBα degradation is associated with macroautophagy (A) NCI-H157 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for Lamp2a, Lamp2, LC3B, and GAPDH. (B) Cells were stimulated with PS-341 (50 nM) for the indicated times. Total RNA was isolated and quantitative real-time PCR for LC3B and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Western Blot, Isolation, Real-time Polymerase Chain Reaction

Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.
Figure Legend Snippet: Macroautophagy mediates PI-induced IκBα degradation (A, B) NCI-H157 cells were transiently transfected with control shRNAs, Lamp2 shRNAs, control siRNAs, or LC3B siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) or MG132 (20 μM) for 8 h. Total cellular extracts were subjected to Western blot analysis for IκBα, Lamp2a, Lamp2, LC3B, and GAPDH. Results are representative of three independent experiments.

Techniques Used: Transfection, Western Blot

PI activates IκB/NF-κB pathway (A) NCI-H157 and A549 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for IκBα and GAPDH. (B) NCI-H157 cells were treated with PS-341 or MG132 for the indicated times. Nuclear proteins were extracted and subjected to Western blot analysis for p65 and Lamin A/C. TNF-α (10 ng/mL, 60 min) was used as a positive control. (C) Both cell lines were stimulated with PS-341 or MG132 for 0, 6, 8, 12, or 24 h. Total RNA was isolated and quantitative real-time PCR for COX-2 and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI activates IκB/NF-κB pathway (A) NCI-H157 and A549 cells were treated with PS-341 (50 nM) or MG132 (20 μM) for the indicated times. Total cellular extracts were subjected to Western blot analysis for IκBα and GAPDH. (B) NCI-H157 cells were treated with PS-341 or MG132 for the indicated times. Nuclear proteins were extracted and subjected to Western blot analysis for p65 and Lamin A/C. TNF-α (10 ng/mL, 60 min) was used as a positive control. (C) Both cell lines were stimulated with PS-341 or MG132 for 0, 6, 8, 12, or 24 h. Total RNA was isolated and quantitative real-time PCR for COX-2 and GAPDH was performed. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Western Blot, Positive Control, Isolation, Real-time Polymerase Chain Reaction

PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P
Figure Legend Snippet: PI-induced Nrf2 activation suppresses PI-induced cell death (A) NCI-H157 cells were transiently transfected with control siRNAs or Nrf2 siRNAs. Forty-eight hours after transfection, the cells were treated with PS-341 (50 nM) for 24 h. Total cellular extracts were subjected to Western blot analysis for PARP, Nrf2, and GAPDH. (B) Cells were infected with adenovirus vectors expressing WT-Nrf2 or DN-Nrf2. After 48 h, the cells were stimulated with PS-341 (50 nM) for 24 h. Cell viability was determined by MTT assay. Data represent the mean ± SD of triplicate experiments. ** P

Techniques Used: Activation Assay, Transfection, Western Blot, Infection, Expressing, MTT Assay

35) Product Images from "Bovine Herpesvirus 1 Productive Infection Led to Inactivation of Nrf2 Signaling through Diverse Approaches"

Article Title: Bovine Herpesvirus 1 Productive Infection Led to Inactivation of Nrf2 Signaling through Diverse Approaches

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2019/4957878

BoHV-1 infection altered the localization of nuclear Nrf2 protein. MDBK cells in 2-well chamber slides were mock infected (a) or infected with BoHV-1 (MOI = 0.1) (b) for 24 hours. After three washings with PBS, cells were fixed with 4% formaldehyde, and Nrf2 was detected by IFA using the Nrf2 antibody (1 : 500) and LaminA/C antibody (1 : 500). DAPI staining was used to stain nuclear DNA. Images were obtained by performing confocal microscopy (Leica). These images are representative of three independent experiments. (c) Zoom-in cells showing typical dot-like staining. (d) The percentage of dot-like staining-positive cells among ~400 cells was estimated from photos derived from three independent experiments. Scale bar: 200 μ M.
Figure Legend Snippet: BoHV-1 infection altered the localization of nuclear Nrf2 protein. MDBK cells in 2-well chamber slides were mock infected (a) or infected with BoHV-1 (MOI = 0.1) (b) for 24 hours. After three washings with PBS, cells were fixed with 4% formaldehyde, and Nrf2 was detected by IFA using the Nrf2 antibody (1 : 500) and LaminA/C antibody (1 : 500). DAPI staining was used to stain nuclear DNA. Images were obtained by performing confocal microscopy (Leica). These images are representative of three independent experiments. (c) Zoom-in cells showing typical dot-like staining. (d) The percentage of dot-like staining-positive cells among ~400 cells was estimated from photos derived from three independent experiments. Scale bar: 200 μ M.

Techniques Used: Infection, Immunofluorescence, Staining, Confocal Microscopy, Derivative Assay

BoHV-1 infection altered the accumulation of Nrf2 in the nucleus. (a) MDBK cells in 60 mm dishes were mock infected or infected with BoHV-1 (MOI = 0.1) for 24 h, the cell cultures were collected for extracting nuclear proteins using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027). Nrf2 was detected by Western blot using the antibody against Nrf2 (Abcam, cat# ab137550). LaminA/C was detected and used as protein loading control. (b) Protein fractions of both the cytosol and nucleus were subjected to Western blots using the antibody against β -tubulin. (c) MDBK cells in 60 mm dishes were mock treated or treated with Trolox (1 mM) for 2 h; the cell lysates were collected for the purification of nuclear proteins and cytosol proteins using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027). Nrf2 was detected by Western blot using the antibody against Nrf2 (Abcam, cat# ab137550). LaminA/C and β -tubulin were detected to characterize whether each fraction was contaminated. These images are representative of three independent experiments.
Figure Legend Snippet: BoHV-1 infection altered the accumulation of Nrf2 in the nucleus. (a) MDBK cells in 60 mm dishes were mock infected or infected with BoHV-1 (MOI = 0.1) for 24 h, the cell cultures were collected for extracting nuclear proteins using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027). Nrf2 was detected by Western blot using the antibody against Nrf2 (Abcam, cat# ab137550). LaminA/C was detected and used as protein loading control. (b) Protein fractions of both the cytosol and nucleus were subjected to Western blots using the antibody against β -tubulin. (c) MDBK cells in 60 mm dishes were mock treated or treated with Trolox (1 mM) for 2 h; the cell lysates were collected for the purification of nuclear proteins and cytosol proteins using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027). Nrf2 was detected by Western blot using the antibody against Nrf2 (Abcam, cat# ab137550). LaminA/C and β -tubulin were detected to characterize whether each fraction was contaminated. These images are representative of three independent experiments.

Techniques Used: Infection, Protein Purification, Western Blot, Purification

BoHV-1 affected the association between Nrf2 and LaminA/C. MDBK cells in 60 mm dishes were mock infected or infected with BoHV-1 for 24 h. Nucleus proteins were purified using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027). IP was performed using antibodies against Nrf2 (a), LaminA/C (b), Nrf2 and unrelated IgG (c), and LaminA/C and IgG (d) (R: rabbit; M: mouse). Then, Western blots were performed using corresponding antibodies. (e) MDBK cells in 60 mm dishes were mock treated or treated with Trolox (1 mM) for 2 h. Nucleus proteins were purified using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027), and LaminA/C was detected by Western blots.
Figure Legend Snippet: BoHV-1 affected the association between Nrf2 and LaminA/C. MDBK cells in 60 mm dishes were mock infected or infected with BoHV-1 for 24 h. Nucleus proteins were purified using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027). IP was performed using antibodies against Nrf2 (a), LaminA/C (b), Nrf2 and unrelated IgG (c), and LaminA/C and IgG (d) (R: rabbit; M: mouse). Then, Western blots were performed using corresponding antibodies. (e) MDBK cells in 60 mm dishes were mock treated or treated with Trolox (1 mM) for 2 h. Nucleus proteins were purified using commercial nuclear protein purification kit (Beyotime Biotechnology, cat# P0027), and LaminA/C was detected by Western blots.

Techniques Used: Infection, Purification, Protein Purification, Western Blot

36) Product Images from "miR-200a Regulates Nrf2 Activation by Targeting Keap1 mRNA in Breast Cancer Cells *"

Article Title: miR-200a Regulates Nrf2 Activation by Targeting Keap1 mRNA in Breast Cancer Cells *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M111.275495

miR-200a promotes Nrf2 activation by targeting Keap1 mRNA. A , miR-200a targets Keap1 mRNA, resulting in Keap1 mRNA destabilization. The Keap1 mRNA stability assay was performed in MDA-MB-231 and Hs578T cell lines transfected with miR-200a expression vector
Figure Legend Snippet: miR-200a promotes Nrf2 activation by targeting Keap1 mRNA. A , miR-200a targets Keap1 mRNA, resulting in Keap1 mRNA destabilization. The Keap1 mRNA stability assay was performed in MDA-MB-231 and Hs578T cell lines transfected with miR-200a expression vector

Techniques Used: Activation Assay, Stability Assay, Multiple Displacement Amplification, Transfection, Expressing, Plasmid Preparation

miR-200a targets Keap1 in breast cancer cell lines. A , array profiling of 88 miRs comparing expression in MCF-10A and MDA-MB-231 cell lines. The scatter plot shows -fold change in miR expression between cell lines (log 10 (2 Δ Ct )). The middle line
Figure Legend Snippet: miR-200a targets Keap1 in breast cancer cell lines. A , array profiling of 88 miRs comparing expression in MCF-10A and MDA-MB-231 cell lines. The scatter plot shows -fold change in miR expression between cell lines (log 10 (2 Δ Ct )). The middle line

Techniques Used: Expressing, Multiple Displacement Amplification

Treatment with SAHA reduces anchorage-independent cell growth in breast cancer cell lines and impacts miR-200a/Keap1/Nrf2 pathway in vivo . A , Western blotting showing Nrf2 protein expression in Nrf2-myc vector- or control vector-transfected MDA-MB-231
Figure Legend Snippet: Treatment with SAHA reduces anchorage-independent cell growth in breast cancer cell lines and impacts miR-200a/Keap1/Nrf2 pathway in vivo . A , Western blotting showing Nrf2 protein expression in Nrf2-myc vector- or control vector-transfected MDA-MB-231

Techniques Used: In Vivo, Western Blot, Expressing, Plasmid Preparation, Transfection, Multiple Displacement Amplification

Epigenetic therapy restores Nrf2 activity in breast cancer cell lines through miR-200a re-expression and Keap1 down-regulation. A , SAHA treatment of breast cancer cell lines results in miR-200a re-expression. miR-200a levels in SAHA-treated Hs578T cells
Figure Legend Snippet: Epigenetic therapy restores Nrf2 activity in breast cancer cell lines through miR-200a re-expression and Keap1 down-regulation. A , SAHA treatment of breast cancer cell lines results in miR-200a re-expression. miR-200a levels in SAHA-treated Hs578T cells

Techniques Used: Activity Assay, Expressing

37) Product Images from "MicroRNA-7450 regulates non-thermal plasma-induced chicken Sertoli cell apoptosis via adenosine monophosphate-activated protein kinase activation"

Article Title: MicroRNA-7450 regulates non-thermal plasma-induced chicken Sertoli cell apoptosis via adenosine monophosphate-activated protein kinase activation

Journal: Scientific Reports

doi: 10.1038/s41598-018-27123-8

ROS production and antioxidant activity of plasma-treated SCs. Chicken SCs were exposed to 22.0 kV of non-thermal plasma for 120 s. ( A ) Imaging of SCs stained with DCFDA/MitoSOX Red/DAPI. Intracellular ROS production was detected by DCFDA staining and mitochondrial superoxide was detected by MitoSOX Red staining. DCFDA: green fluorescence; MitoSOX Red: red fluorescence; DAPI: blue fluorescence. Scale bar: 50 μm. ( B ) Relative fluorescence intensity for DCFDA staining. ( C ) Relative fluorescence intensity for MitoSOX Red staining. ( D ) Total ROS levels in SCs. ( E ) MDA level in SCs. Activities of ( F ) SOD, ( G ) CAT, and ( H ) GPx in SCs. Relative mRNA levels of ( I ) NOX4 , NRF2 , and KEAP1 , and ( J ) SOD , CAT , GPx , and PRDX4 . Data are represented as the mean ± SD (n = 3 per group). * p
Figure Legend Snippet: ROS production and antioxidant activity of plasma-treated SCs. Chicken SCs were exposed to 22.0 kV of non-thermal plasma for 120 s. ( A ) Imaging of SCs stained with DCFDA/MitoSOX Red/DAPI. Intracellular ROS production was detected by DCFDA staining and mitochondrial superoxide was detected by MitoSOX Red staining. DCFDA: green fluorescence; MitoSOX Red: red fluorescence; DAPI: blue fluorescence. Scale bar: 50 μm. ( B ) Relative fluorescence intensity for DCFDA staining. ( C ) Relative fluorescence intensity for MitoSOX Red staining. ( D ) Total ROS levels in SCs. ( E ) MDA level in SCs. Activities of ( F ) SOD, ( G ) CAT, and ( H ) GPx in SCs. Relative mRNA levels of ( I ) NOX4 , NRF2 , and KEAP1 , and ( J ) SOD , CAT , GPx , and PRDX4 . Data are represented as the mean ± SD (n = 3 per group). * p

Techniques Used: Antioxidant Activity Assay, Imaging, Staining, Fluorescence, Multiple Displacement Amplification

Chicken SC protein expression. ( A . Relative protein levels of ( B ) NRF2, KEAP1, PRDX4, ( C ) ATP5A, ( D ) p-AMPKα/AMPKα, and ( E ) p-mTOR/mTOR in SCs exposed to plasma. ( F . Relative protein levels of ( G ) ATP5A, ( H ) p-AMPKα/AMPKα, and ( I . Data are represented as the mean ± SD (n = 3 per group). * p
Figure Legend Snippet: Chicken SC protein expression. ( A . Relative protein levels of ( B ) NRF2, KEAP1, PRDX4, ( C ) ATP5A, ( D ) p-AMPKα/AMPKα, and ( E ) p-mTOR/mTOR in SCs exposed to plasma. ( F . Relative protein levels of ( G ) ATP5A, ( H ) p-AMPKα/AMPKα, and ( I . Data are represented as the mean ± SD (n = 3 per group). * p

Techniques Used: Expressing

38) Product Images from "Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling"

Article Title: Blockade of RAGE ameliorates elastase-induced emphysema development and progression via RAGE-DAMP signaling

Journal: The FASEB Journal

doi: 10.1096/fj.201601155R

Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD. A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis. B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin. C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P
Figure Legend Snippet: Effect of FPS-ZM1 on activation of DAMP signaling in PPE-induced experimental COPD. A ) IKKα, IKKβ, and phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) were determined by Western blot analysis. B ) JNK/ERK/p38 MAPK activation (phosphorylation) was determined by Western blot with the appropriate antibodies. The target protein bands were subjected to densitometric analysis normalized with β-actin. C ) The canonical–noncanonical Rel/p65 NF-κB activation and MAPK activation and its correlation to Nrf2 translocation from the cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for the nuclear protein loading control, and β-actin was used for the cytoplasmic protein loading control. The corresponding density ratio was determined by the average intensity of the bands. Ctl, control group; ns, not significant. * P

Techniques Used: Activation Assay, Western Blot, Translocation Assay, CTL Assay

Effect of FPS-ZM1 on CSE-induced activation of DAMP signaling in MLE-12 cells. A – C ) After exposure to CSE in the presence or absence of FPS-ZM1 for 24 h, Western blot analysis was perfomed ( A ), and the expression levels of mRAGE in lung homogenates ( B ) and of circulating sRAGE ( C ) were determined by ELISA. D , E ) IKKα, IKKβ, and the phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) ( D ) and the phosphorylation of JNK/ERK/p38 MAPK activation ( E ) were detected with the appropriate antibodies. β-Actin, total RelA/p65, total JNK1/2, total ERK1/2, and total p38 were used for loading controls. F ) Correlation to Nrf2 translocation from cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for nuclear protein loading control, and β-actin was used for cytoplasmic protein loading control. The activation of DAMP signaling was inhibited by FPS-ZM1 treatment. Results are combined data from 3 independent experiments. Ctl, control group; ns, not significant. * P
Figure Legend Snippet: Effect of FPS-ZM1 on CSE-induced activation of DAMP signaling in MLE-12 cells. A – C ) After exposure to CSE in the presence or absence of FPS-ZM1 for 24 h, Western blot analysis was perfomed ( A ), and the expression levels of mRAGE in lung homogenates ( B ) and of circulating sRAGE ( C ) were determined by ELISA. D , E ) IKKα, IKKβ, and the phosphorylation of Rel/p65 at 2 sites (Ser276 and Ser536) ( D ) and the phosphorylation of JNK/ERK/p38 MAPK activation ( E ) were detected with the appropriate antibodies. β-Actin, total RelA/p65, total JNK1/2, total ERK1/2, and total p38 were used for loading controls. F ) Correlation to Nrf2 translocation from cytoplasm into the nucleus was measured by Western blot analysis. Lamin A/C was used for nuclear protein loading control, and β-actin was used for cytoplasmic protein loading control. The activation of DAMP signaling was inhibited by FPS-ZM1 treatment. Results are combined data from 3 independent experiments. Ctl, control group; ns, not significant. * P

Techniques Used: Activation Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Translocation Assay, CTL Assay

39) Product Images from "Schisandrol B protects against acetaminophen-induced acute hepatotoxicity in mice via activation of the NRF2/ARE signaling pathway"

Article Title: Schisandrol B protects against acetaminophen-induced acute hepatotoxicity in mice via activation of the NRF2/ARE signaling pathway

Journal: Acta Pharmacologica Sinica

doi: 10.1038/aps.2015.120

The effects of SolB on activation and expression of NRF2 in vivo . (A) qRT-PCR analysis was performed to measure the gene expression of Nrf2, and the data are expressed as the mean±SEM ( n =6). (B–C) Western blotting was used to measure nuclear
Figure Legend Snippet: The effects of SolB on activation and expression of NRF2 in vivo . (A) qRT-PCR analysis was performed to measure the gene expression of Nrf2, and the data are expressed as the mean±SEM ( n =6). (B–C) Western blotting was used to measure nuclear

Techniques Used: Activation Assay, Expressing, In Vivo, Quantitative RT-PCR, Western Blot

Role of NRF2 signaling and its downstream proteins in SolB-mediated protection against APAP-induced liver toxicity. (A) Western blotting was used to measure the protein expression of GSR, GSS, GCLC, GCLM, GST-α, GST-μ, GST-π, NQO1,
Figure Legend Snippet: Role of NRF2 signaling and its downstream proteins in SolB-mediated protection against APAP-induced liver toxicity. (A) Western blotting was used to measure the protein expression of GSR, GSS, GCLC, GCLM, GST-α, GST-μ, GST-π, NQO1,

Techniques Used: Western Blot, Expressing

Effects of SolB on cell viability and NRF2 activation. (A) An MTT assay was used to measure HepG2 cell viability after treatment with SolB at 2.5 to 160 μmol/L. (B) A luciferase reporter assay was used to measure the effect of SolB on NRF2 activation.
Figure Legend Snippet: Effects of SolB on cell viability and NRF2 activation. (A) An MTT assay was used to measure HepG2 cell viability after treatment with SolB at 2.5 to 160 μmol/L. (B) A luciferase reporter assay was used to measure the effect of SolB on NRF2 activation.

Techniques Used: Activation Assay, MTT Assay, Luciferase, Reporter Assay

40) Product Images from "Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity"

Article Title: Sulforaphane Protects the Liver against CdSe Quantum Dot-Induced Cytotoxicity

Journal: PLoS ONE

doi: 10.1371/journal.pone.0138771

Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P
Figure Legend Snippet: Effect of knockdown TR-1, Keap1 and Nrf2 on cytotoxicity in HHL-5 cells exposed to CdSe QDs. TR-1, Keap1 or Nrf2 were knocked down, respectively. Allstars (AS) were used as a negative control. Cells were incubated with 5 μM SFN or DMSO (0.1%) control for 24 h then exposed to 20 μM CdSe QDs for 24 h. Cell viability was measured by MTT assay, and data shown as means ± SD (n = 6). Significant levels (*P

Techniques Used: Negative Control, Incubation, MTT Assay

Related Articles

Transduction:

Article Title: A Role for Nrf2 Expression in Defining the Aging of Hippocampal Neural Stem Cells
Article Snippet: .. Moreover, the number of cells in Nrf2 overexpressing middle-aged grafts was also higher (although not significantly, p > 0.05) compared with control middle-aged grafts transduced with just eGFP ( ). ..

Transfection:

Article Title: A Role for Nrf2 Expression in Defining the Aging of Hippocampal Neural Stem Cells
Article Snippet: .. Under these conditions, interestingly the survival of the cells ( ) was not significantly affected however, the proliferation substantially improved ( , p < 0.001, untreated versus Nrf2 transfected). .. We additionally also assessed DG NSPCs from newborn (postnatal day 0) Nrf2 knockout (Nrf2-/-) and WT (Nrf2+/+) mice.

other:

Article Title: A Role for Nrf2 Expression in Defining the Aging of Hippocampal Neural Stem Cells
Article Snippet: Finally, in the pattern separation test, the Nrf2-/- animals exhibited a compromised behavior as indicated by their substantially reduced exploration of the object in the novel context (p < 0.05, ) when compared with their WT counterparts.

Article Title: Antioxidant Properties of Fullerene Derivatives Depend on Their Chemical Structure: A Study of Two Fullerene Derivatives on HELFs
Article Snippet: NRF2 (erythroid-derived factor 2) is one of the main transcription factors that determine antioxidant response of the cells to the action of the internal and external ROS.

Article Title: Nrf2-mediated anti-oxidant effects contribute to suppression of non-alcoholic steatohepatitis-associated hepatocellular carcinoma in murine model
Article Snippet: The results of this study support that Nrf2 and its related metabolites have protective effects on liver injury, inflammation, and tumorigenesis. ( , )

Article Title: Antioxidant Properties of Fullerene Derivatives Depend on Their Chemical Structure: A Study of Two Fullerene Derivatives on HELFs
Article Snippet: After 3 h with 4 nM of fullerene, the amount of NRF2 is reduced.

Article Title: Lipin1 deficiency causes sarcoplasmic reticulum stress and chaperone‐responsive myopathy
Article Snippet: Reagents The following primary antibodies were used: anti‐p62 (SQSTM) (Abnova), anti‐LAMP2 (Abcam), anti‐FGF21 (abcam), anti‐Bip (BD Biosciences), anti‐Gapdh (Santa Cruz), anti‐SREBP1c (Santa Cruz), anti‐SREBP2 (abcam), anti‐ATF6 (abcam), anti‐LC3 (Nanotools), anti‐Tom20 (SantaCruz), anti‐lipin1 (SantaCruz, sc‐376874).

Expressing:

Article Title: A Role for Nrf2 Expression in Defining the Aging of Hippocampal Neural Stem Cells
Article Snippet: .. Here, a significant reduction in MCM2 , Sox2 , and GFAP/nestin ( ) expressing NSPCs was noted in the DG of the Nrf2-/- mice. .. Moreover, the number of Dcx+ newborn neurons was also significantly reduced in the Nrf2-/- mice compared with WT controls ( ).

Mouse Assay:

Article Title: A Role for Nrf2 Expression in Defining the Aging of Hippocampal Neural Stem Cells
Article Snippet: .. Here, a significant reduction in MCM2 , Sox2 , and GFAP/nestin ( ) expressing NSPCs was noted in the DG of the Nrf2-/- mice. .. Moreover, the number of Dcx+ newborn neurons was also significantly reduced in the Nrf2-/- mice compared with WT controls ( ).

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    Santa Cruz Biotechnology anti keap1
    Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and <t>Keap1</t> proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p
    Anti Keap1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and <t>Keap1</t> proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p
    Goat Anti Keap1 Polyclonal Antibody E20, 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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    Santa Cruz Biotechnology goat polyclonal anti kelch like ech associated protein 1 keap1 antibody
    Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and <t>Keap1</t> proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p
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    Image Search Results


    Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and Keap1 proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p

    Journal: Molecules (Basel, Switzerland)

    Article Title: The Cinnamon-derived Dietary Factor Cinnamic Aldehyde Activates the Nrf2-dependent Antioxidant Response in Human Epithelial Colon Cells

    doi: 10.3390/molecules15053338

    Figure Lengend Snippet: Dose response of Nrf2 transcriptional activation by cinnamic aldehyde and an ethanolic cinnamon extract in MDA-MB231 breast carcinoma and HCT116 colon carcinoma cells. (a–b) MDA-MB-231 cells (panel a) or HCT116 (panel b) were cotransfected with plasmids containing a GST-ARE-firefly luciferase reporter gene and expression plasmids for the Nrf2 and Keap1 proteins. A plasmid encoding renilla luciferase, driven by the herpes simplex virus thymidine kinase promoter, was included in all transfections to normalize transfection efficiency. Twenty-four hours post-transfection, cells were dosed with the indicated concentrations of each compound (tBHQ: 50μM) for 16h. Firefly and renilla luciferase activity was measured and is expressed as relative activity (F/R) compared to untreated control ( p

    Article Snippet: Western blot analysis of Nrf2, Keap1, γ-GCS and HO-1 was performed as published recently using anti-Nrf2 (H-300, rabbit polyclonal), anti-Keap1 (E-20, goat polyclonal) anti-GCSm (E-4, mouse monoclonal), and anti-heme oxygenase 1 (H-105, rabbit polyclonal) antibodies, respectively (Santa Cruz Biotechnology, Santa Cruz, CA) [ – ].

    Techniques: Activation Assay, Multiple Displacement Amplification, Luciferase, Expressing, Plasmid Preparation, Transfection, Activity Assay