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  • 94
    Millipore nqo 1
    (A) The protein levels of nuclear transcription factor NF-E2-related factor 2 (Nrf-2), (B) heme oxygenase-1 (HO-1) and (C) NAD(P)H:quinone oxidoreductase-1 enzyme <t>(NQO-1)</t> were compared using western blot analysis of lung tissue extracts at 15, 30, 60 and 20 days post-irradiation. The relative protein expression levels of Nrf-2, HO-1, and NQO-1 in lung homogenates were significantly higher (p
    Nqo 1, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    ABclonal nqo 1
    Oroxylin A-driven p62 cleavage down-regulated Nrf2 to induce oxidative stress. (A) Domain composition of human p62/SQSTM1 showing the caspase-8 cleavage site (not to scale). Blue arrows point to regions for interactions with the indicated proteins. (B) The IL-1β and IL-6 mRNA was measured by real-time PCR following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. (C) The HO-1, SOD1 and <t>NQO-1</t> mRNA was measured by real-time PCR following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. β-actin was used as internal control. The relative levels were calculated to β-actin mRNA expression. (D) The levels of Nrf2, HO-1 and NQO-1 were assessed by western blot following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. (E) The ROS level was assessed following 64 μM oroxylin A and 50 μM Z-IETD-FMK treatment in SMMC-7721 cells. Data are presented as mean ± SD. *P
    Nqo 1, supplied by ABclonal, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Novus Biologicals nqo1
    Effect of leptin on <t>NQO1</t> in MCF-7 cells.
    Nqo1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 91/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nqo 1  (Abcam)
    94
    Abcam nqo 1
    Resveratrol strengthens activation of the Nrf-2 signaling pathway at 24 h after OGD/R injury in vitro. (A) Neurons were immunostained with antibodies to Nrf-2 (green). Nuclei were labeled with PI (red). Nrf-2 was mainly located in the cytoplasm in the normal group (A,left) . There were a few cells positive for Nrf-2 in the nuclei in the model group (A,middle). Nrf-2 was mainly located in the nucleus in the resveratrol group (A,right). (B) Protein expressions of Nrf-2, HO-1, and <t>NQO-1</t> with western blot analysis. (C,D) Quantification of data for Nrf2, NQO-1, and HO-1 proteins. The protein expression levels of Nrf2 in the nuclei, and NQO-1 and HO-1 in the cytoplasm, were significantly upregulated in the Mod and Res groups compared to the Nor group, and were highest in the WP group. This suggests that resveratrol increased expression of Nrf2 in the nuclei and NQO-1 and HO-1 in the cytoplasm. *P
    Nqo 1, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 190 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Novus Biologicals rabbit anti nqo1
    Defective autophagy in macrophages triggers neither an antioxidative backup mechanism nor senescence. Bone marrow-derived macrophages were isolated from Atg7 +/+ Lysm-Cre + (+/+) and Atg7 F/F Lysm-Cre + (−/−) mice. ( A ) Western blot analysis of ATG7 and SQSTM1 in untreated Atg7 +/+ and atg7 −/− macrophages. ( B ) Real-time RT-PCR and western blot analysis of GSTA and <t>NQO1</t> in Atg7 +/+ and atg7 −/− macrophages (NS, not significant; n = 2 experiments in duplicate; Student t test). ( C ) Atg7 +/+ and atg7 −/− macrophages were treated with 50 µmol/l tBHP for 24 h (***, P
    Rabbit Anti Nqo1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam nqo 1 antibodies
    Robo-2 is required for the targeting of MOR174–9-expressing OSN axons in the OB. A–J , Coronal sections of OBs isolated from control ( A , D , G , H ) and robo-2 lox/lox ;OMP-Cre; MOR174–9-tau-GFP ( B , C , E , F , I , J ) mice stained with <t>NQO-1</t> and GFP antibodies ( A–J ). NQO-1 antibodies stain axons of OSNs innervating the dorsal region of the OB. On the lateral side of the OB, a GFP-positive glomerulus is consistently observed in the dorsal region of the OB ( A–C ) and a second GFP-positive glomerulus is observed in approximately half of the OBs analyzed (14/28 controls and 18/25 robo-2 lox/lox ; OMP-Cre ) ( D–F ). In addition, a third glomerulus was observed more ventrally in two OBs from control mice and in six OBs from robo-2 lox/lox ; OMP-Cre mice (shown in yellow on the scatter plot in K ). On the medial side of the OB, all OBs analyzed contained at least one GFP-positive glomerulus ( G , H ) and the majority of OBs analyzed (19/27 for controls and 19/25 for robo-2 lox/lox ; OMP-Cre mice) contained two GFP-positive glomeruli ( H , J ). Furthermore, a few OBs analyzed contained an additional GFP-positive glomerulus (2/28 controls and 6/27 robo-2 lox/lox ; OMP-Cre ). K , L , The relative positions of GFP-positive glomeruli in the dorsoventral axis of the OB on the lateral and medial sides of the OB were assessed and represented on scatter plots ( K , L ). Parameters are shown as mean ± SEM. The mean location of glomeruli innervated by MOR174–9-expressing axons on both the lateral and medial sides of the OB is shifted ventrally in robo-2 lox/lox ; OMP-Cre . Note that the additional glomeruli observed in some bulbs (represented in yellow) were not included in the calculation of the mean. Sections containing glomeruli located close to the mean angle are shown in A , B , D , E , G–J . In C and F , sections with a more severe ventral shift in the location of the glomeruli are shown. *** p
    Nqo 1 Antibodies, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Novus Biologicals nqo1 antibody
    Equol attenuated ER stress by activating Nrf2 in vitro and in vivo. (A) HUVECs were incubated with different concentrations (1, 10 and 100 nM) of equol for 24 h before treatment with t -BHP (50 μM) for another 6 h. The expression of Nrf2 and <t>NQO1</t> was detected by western blotting. (C) Total tissue lysates from the thoracic and abdominal aorta of apoE-/- mice with or without equol treatment were immunoblotted with anti-Nrf2 and anti-β-actin antibodies. (E) Cells were transfected with Nrf2 siRNA for 5~6 h; the medium was then replaced with fresh culture medium, followed by incubation for another 24 h. Thereafter, the cells were treated with equol (100 nM) for 24 h and then were incubated with t -BHP (50 μM) for an additional 6 h. The cells were collected and lysed, and western blot analysis was performed. (B)(D)(F) The bar charts show the quantification of the indicated proteins. (G) HUVECs were transfected with Nrf2 siRNA and treated as described in (C), and cell apoptosis was assayed using the Cell Death Detection ELISA plus Kit. Values are presented as means ± SD. n = 3, * p
    Nqo1 Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Novus Biologicals nqo1 protein
    Expression and activity of <t>Nqo1</t> in liver cytosolic fractions from wild-type and Nrf2-null mice 1 and 3 days after sham or BDL surgery. (A) Total RNA was isolated from livers of WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously, and analyzed by the bDNA assay for Nqo1 mRNA expression. Data from sham animals at 1 and 3 days were not statistically different and were pooled and denoted as sham. The data are presented as mean relative light units ± standard error of the mean (SEM) ( n = 4–7 animals). (B) Upper panel: Representative Western blot of liver cytosolic fractions from mice 3 days after sham or BDL surgery ( n = 4–7 animals) stained with anti-Nqo1 antibodies. Lower panel: Quantification of Nqo1 protein levels in liver cytosolic fractions from mice 1 and 3 days after sham or BDL surgery. The data are presented as relative protein expression ± SEM ( n = 4–7 animals). (C) Analysis of Nqo1 activity in liver cytosolic fractions from WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously. The data are presented as nanomoles of DCPIP reduced per minute per milligram of protein ± SEM ( n = 4–7 animals). Asterisks (*) represent a statistical difference ( P
    Nqo1 Protein, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Novus Biologicals goat anti nqo1 polyclonal antibody
    Expression and activity of <t>Nqo1</t> in liver cytosolic fractions from wild-type and Nrf2-null mice 1 and 3 days after sham or BDL surgery. (A) Total RNA was isolated from livers of WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously, and analyzed by the bDNA assay for Nqo1 mRNA expression. Data from sham animals at 1 and 3 days were not statistically different and were pooled and denoted as sham. The data are presented as mean relative light units ± standard error of the mean (SEM) ( n = 4–7 animals). (B) Upper panel: Representative Western blot of liver cytosolic fractions from mice 3 days after sham or BDL surgery ( n = 4–7 animals) stained with anti-Nqo1 antibodies. Lower panel: Quantification of Nqo1 protein levels in liver cytosolic fractions from mice 1 and 3 days after sham or BDL surgery. The data are presented as relative protein expression ± SEM ( n = 4–7 animals). (C) Analysis of Nqo1 activity in liver cytosolic fractions from WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously. The data are presented as nanomoles of DCPIP reduced per minute per milligram of protein ± SEM ( n = 4–7 animals). Asterisks (*) represent a statistical difference ( P
    Goat Anti Nqo1 Polyclonal Antibody, supplied by Novus Biologicals, 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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    94
    Santa Cruz Biotechnology nqo 1
    Salvianolic acid A (SalA) regulates the protein expression of apoptosis-related proteins, Nrf2 signaling, and MAPK signaling. A: Representative protein bands of Bax, Bcl-2, caspase-3 and caspase-9 expression in control, subarachnoid hemorrhage (SAH), and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. B: Representative protein bands of Nrf2, HO-1 and <t>NQO-1</t> expression in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. C: Representative protein bands of the proteins and the phosphorylation of the proteins in MAPK signaling in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. Values are represented as mean ± standard deviation (n = 5, each group). #P
    Nqo 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Epitomics nqo 1
    Salvianolic acid A (SalA) regulates the protein expression of apoptosis-related proteins, Nrf2 signaling, and MAPK signaling. A: Representative protein bands of Bax, Bcl-2, caspase-3 and caspase-9 expression in control, subarachnoid hemorrhage (SAH), and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. B: Representative protein bands of Nrf2, HO-1 and <t>NQO-1</t> expression in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. C: Representative protein bands of the proteins and the phosphorylation of the proteins in MAPK signaling in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. Values are represented as mean ± standard deviation (n = 5, each group). #P
    Nqo 1, supplied by Epitomics, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cell Signaling Technology Inc nqo 1
    Polydatin protected BMSCs against H 2 O 2 -induced cell death partly through Nrf 2/ARE pathway. BMSCs were pretreated with polydatin for 2 h and further exposed to H 2 O 2 for 12 h. (a) Effects of polydatin on <t>NQO-1</t> and the phosphorylation of Nrf 2. (b, c) Quantitative analysis of the blots was shown in panel after being normalized by α -tubulin. (d) Cells were treated with different concentration of brusatol for 24 h. Effects of brusatol on phosphorylation of Nrf 2 were detected by Western blot and (g) the bands were normalized by α -tubulin. (e) Cell viability was tested in the presence of different concentration of brusatol. (h) BMSCs were pretreated with brusatol (100 μ M) for 1 h followed by incubating with/without polydatin and H 2 O 2 for 24 h. (f) ROS production was detected by H2DCF-DA staining. (b) Quantitative analysis of DCF fluorescent intensity. One-way ANOVA followed by Tukey's test. ∗ p
    Nqo 1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Novus Biologicals anti nqo1
    Antioxidant enzyme synthesis in response to orange oil treatment . Immunoblot analysis demonstrating expression of HO-1, <t>NQO1</t> and GCLm proteins at 6, 12 and 24 hrs following 15 min treatment of BEAS-2B cells with the oil preparation or time-matched soy oil control. Representative blots from one of three separate experiments are shown above. Densitometric evaluations of each target protein blot normalized to its corresponding GAPDH for all three experiments are provided below. Bars represent mean ± SEM.
    Anti Nqo1, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    R&D Systems human nqo 1 antibody
    Antioxidant enzyme synthesis in response to orange oil treatment . Immunoblot analysis demonstrating expression of HO-1, <t>NQO1</t> and GCLm proteins at 6, 12 and 24 hrs following 15 min treatment of BEAS-2B cells with the oil preparation or time-matched soy oil control. Representative blots from one of three separate experiments are shown above. Densitometric evaluations of each target protein blot normalized to its corresponding GAPDH for all three experiments are provided below. Bars represent mean ± SEM.
    Human Nqo 1 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Proteintech nqo 1
    Effect of MitoQ on the Nrf2 signalling in compression‐exposed human NP cells. (A‐J) Human NP cells were treated with or without MitoQ (500 nmol/L) for 12 h. (A‐C) The protein levels of Nrf2 and Keap1 in the human NP cells were measured by Western blotting. (D‐F) The protein expression of nucleus Nrf2 (Nu‐Nrf2) and cytoplasmic Nrf2 (Cyto‐Nrf2) was measured by Western blotting. (G) The nuclear translocation of Nrf2 was examined by immunofluorescence on confocal microscope. Scale bar: 10 μm. (H‐K) The protein levels of SOD‐2, HO‐1 and <t>NQO‐1</t> in the human NP cells were measured by Western blotting. (L‐V) Human NP cells were pre‐treated with MitoQ (500 nmol/L) for 2 h and then exposed to compression for 36 h. (L‐Q) The protein levels of Nrf2, Keap1, Nu‐Nrf2 and cyto‐Nrf2 in the human NP cells were measured by Western blotting. (R) The nuclear translocation of Nrf2 was examined by immunofluorescence on confocal microscope. Scale bar: 10 μm. (S‐V) The protein levels of SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. Data are represented as the mean ± SD. *** P
    Nqo 1, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher nqo 1
    Effects of C66 on aortic Nrf2 function. Aortic expression of p-Nrf2 was examined by immunohistochemical staining (A), followed semi-quantitative analysis (B), and Nrf2-downstream genes CAT (C) and <t>NQO-1</t> (D) expression was examined by real-time PCR at mRNA level. Data were presented as means ± SD ( n = 5); * P
    Nqo 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 75 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam polyclonal goat anti nqo 1
    Effects of C66 on aortic Nrf2 function. Aortic expression of p-Nrf2 was examined by immunohistochemical staining (A), followed semi-quantitative analysis (B), and Nrf2-downstream genes CAT (C) and <t>NQO-1</t> (D) expression was examined by real-time PCR at mRNA level. Data were presented as means ± SD ( n = 5); * P
    Polyclonal Goat Anti Nqo 1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    (A) The protein levels of nuclear transcription factor NF-E2-related factor 2 (Nrf-2), (B) heme oxygenase-1 (HO-1) and (C) NAD(P)H:quinone oxidoreductase-1 enzyme (NQO-1) were compared using western blot analysis of lung tissue extracts at 15, 30, 60 and 20 days post-irradiation. The relative protein expression levels of Nrf-2, HO-1, and NQO-1 in lung homogenates were significantly higher (p

    Journal: International Journal of Molecular Medicine

    Article Title: The green tea extract epigallocatechin-3-gallate inhibits irradiation-induced pulmonary fibrosis in adult rats

    doi: 10.3892/ijmm.2014.1745

    Figure Lengend Snippet: (A) The protein levels of nuclear transcription factor NF-E2-related factor 2 (Nrf-2), (B) heme oxygenase-1 (HO-1) and (C) NAD(P)H:quinone oxidoreductase-1 enzyme (NQO-1) were compared using western blot analysis of lung tissue extracts at 15, 30, 60 and 20 days post-irradiation. The relative protein expression levels of Nrf-2, HO-1, and NQO-1 in lung homogenates were significantly higher (p

    Article Snippet: The membranes were then incubated with the following primary antibodies at room temperature for 3 h: Nrf-2 (1:200; Sigma), HO-1 (1:200), NQO-1 (1:200) (both from Millipore) or β-actin (1:2,000; Cell Signaling Technology, Inc., Danvers, MA, USA).

    Techniques: Western Blot, Irradiation, Expressing

    (A) Epigallocatechin-3-gallate (EGCG) activates nuclear transcription factor NF-E2-related factor 2 (Nrf-2), (B) heme oxygenase-1 (HO-1) and (C) NAD(P)H:quinone oxidoreductase-1 (NQO-1) protein expression as detected by western blot analysis of lung tissue extracts at 15 days post-irradiation. Immunoblot analysis revealed that the protein expression of Nrf-2, HO-1, and NQO-1 was strongly activated by EGCG administration, while Nrf-2 was weakly activated by dexamethasone (DEX).

    Journal: International Journal of Molecular Medicine

    Article Title: The green tea extract epigallocatechin-3-gallate inhibits irradiation-induced pulmonary fibrosis in adult rats

    doi: 10.3892/ijmm.2014.1745

    Figure Lengend Snippet: (A) Epigallocatechin-3-gallate (EGCG) activates nuclear transcription factor NF-E2-related factor 2 (Nrf-2), (B) heme oxygenase-1 (HO-1) and (C) NAD(P)H:quinone oxidoreductase-1 (NQO-1) protein expression as detected by western blot analysis of lung tissue extracts at 15 days post-irradiation. Immunoblot analysis revealed that the protein expression of Nrf-2, HO-1, and NQO-1 was strongly activated by EGCG administration, while Nrf-2 was weakly activated by dexamethasone (DEX).

    Article Snippet: The membranes were then incubated with the following primary antibodies at room temperature for 3 h: Nrf-2 (1:200; Sigma), HO-1 (1:200), NQO-1 (1:200) (both from Millipore) or β-actin (1:2,000; Cell Signaling Technology, Inc., Danvers, MA, USA).

    Techniques: Expressing, Western Blot, Irradiation

    Oroxylin A-driven p62 cleavage down-regulated Nrf2 to induce oxidative stress. (A) Domain composition of human p62/SQSTM1 showing the caspase-8 cleavage site (not to scale). Blue arrows point to regions for interactions with the indicated proteins. (B) The IL-1β and IL-6 mRNA was measured by real-time PCR following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. (C) The HO-1, SOD1 and NQO-1 mRNA was measured by real-time PCR following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. β-actin was used as internal control. The relative levels were calculated to β-actin mRNA expression. (D) The levels of Nrf2, HO-1 and NQO-1 were assessed by western blot following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. (E) The ROS level was assessed following 64 μM oroxylin A and 50 μM Z-IETD-FMK treatment in SMMC-7721 cells. Data are presented as mean ± SD. *P

    Journal: Redox Biology

    Article Title: Triggering apoptosis by oroxylin A through caspase-8 activation and p62/SQSTM1 proteolysis

    doi: 10.1016/j.redox.2019.101392

    Figure Lengend Snippet: Oroxylin A-driven p62 cleavage down-regulated Nrf2 to induce oxidative stress. (A) Domain composition of human p62/SQSTM1 showing the caspase-8 cleavage site (not to scale). Blue arrows point to regions for interactions with the indicated proteins. (B) The IL-1β and IL-6 mRNA was measured by real-time PCR following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. (C) The HO-1, SOD1 and NQO-1 mRNA was measured by real-time PCR following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. β-actin was used as internal control. The relative levels were calculated to β-actin mRNA expression. (D) The levels of Nrf2, HO-1 and NQO-1 were assessed by western blot following 64 μM oroxylin A treatment or p62 knockdown in SMMC-7721 cells. (E) The ROS level was assessed following 64 μM oroxylin A and 50 μM Z-IETD-FMK treatment in SMMC-7721 cells. Data are presented as mean ± SD. *P

    Article Snippet: Primary antibodies against to caspase-8, caspase-9, cleaved-caspase-3, PARP, FADD, NQO-1 and β-actin were obtained from ABclonal (ABclonal, Wuhan, China).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot

    Effect of leptin on NQO1 in MCF-7 cells.

    Journal: Cancer Biology & Medicine

    Article Title: Leptin influences estrogen metabolism and increases DNA adduct formation in breast cancer cells

    doi: 10.20892/j.issn.2095-3941.2016.0079

    Figure Lengend Snippet: Effect of leptin on NQO1 in MCF-7 cells.

    Article Snippet: In the Reporter assay, leptin (100 ng/mL) significantly downregulated NQO1 from 100% to 45.78% ( ).

    Techniques:

    Resveratrol strengthens activation of the Nrf-2 signaling pathway at 24 h after OGD/R injury in vitro. (A) Neurons were immunostained with antibodies to Nrf-2 (green). Nuclei were labeled with PI (red). Nrf-2 was mainly located in the cytoplasm in the normal group (A,left) . There were a few cells positive for Nrf-2 in the nuclei in the model group (A,middle). Nrf-2 was mainly located in the nucleus in the resveratrol group (A,right). (B) Protein expressions of Nrf-2, HO-1, and NQO-1 with western blot analysis. (C,D) Quantification of data for Nrf2, NQO-1, and HO-1 proteins. The protein expression levels of Nrf2 in the nuclei, and NQO-1 and HO-1 in the cytoplasm, were significantly upregulated in the Mod and Res groups compared to the Nor group, and were highest in the WP group. This suggests that resveratrol increased expression of Nrf2 in the nuclei and NQO-1 and HO-1 in the cytoplasm. *P

    Journal: Cell Transplantation

    Article Title: Resveratrol Treatment in Different Time-Attenuated Neuronal Apoptosis After Oxygen and Glucose Deprivation/Reoxygenation via Enhancing the Activation of Nrf-2 Signaling Pathway In Vitro

    doi: 10.1177/0963689718780930

    Figure Lengend Snippet: Resveratrol strengthens activation of the Nrf-2 signaling pathway at 24 h after OGD/R injury in vitro. (A) Neurons were immunostained with antibodies to Nrf-2 (green). Nuclei were labeled with PI (red). Nrf-2 was mainly located in the cytoplasm in the normal group (A,left) . There were a few cells positive for Nrf-2 in the nuclei in the model group (A,middle). Nrf-2 was mainly located in the nucleus in the resveratrol group (A,right). (B) Protein expressions of Nrf-2, HO-1, and NQO-1 with western blot analysis. (C,D) Quantification of data for Nrf2, NQO-1, and HO-1 proteins. The protein expression levels of Nrf2 in the nuclei, and NQO-1 and HO-1 in the cytoplasm, were significantly upregulated in the Mod and Res groups compared to the Nor group, and were highest in the WP group. This suggests that resveratrol increased expression of Nrf2 in the nuclei and NQO-1 and HO-1 in the cytoplasm. *P

    Article Snippet: Western blot analysis showed that the protein levels of Nrf-2 in the nucleus, and NQO-1 and HO-1 in the cytoplasm, were significantly increased in the model and resveratrol groups compared to those in the normal group ( P < 0.05).

    Techniques: Activation Assay, In Vitro, Labeling, Western Blot, Expressing

    Defective autophagy in macrophages triggers neither an antioxidative backup mechanism nor senescence. Bone marrow-derived macrophages were isolated from Atg7 +/+ Lysm-Cre + (+/+) and Atg7 F/F Lysm-Cre + (−/−) mice. ( A ) Western blot analysis of ATG7 and SQSTM1 in untreated Atg7 +/+ and atg7 −/− macrophages. ( B ) Real-time RT-PCR and western blot analysis of GSTA and NQO1 in Atg7 +/+ and atg7 −/− macrophages (NS, not significant; n = 2 experiments in duplicate; Student t test). ( C ) Atg7 +/+ and atg7 −/− macrophages were treated with 50 µmol/l tBHP for 24 h (***, P

    Journal: Autophagy

    Article Title: Defective autophagy in vascular smooth muscle cells accelerates senescence and promotes neointima formation and atherogenesis

    doi: 10.1080/15548627.2015.1096485

    Figure Lengend Snippet: Defective autophagy in macrophages triggers neither an antioxidative backup mechanism nor senescence. Bone marrow-derived macrophages were isolated from Atg7 +/+ Lysm-Cre + (+/+) and Atg7 F/F Lysm-Cre + (−/−) mice. ( A ) Western blot analysis of ATG7 and SQSTM1 in untreated Atg7 +/+ and atg7 −/− macrophages. ( B ) Real-time RT-PCR and western blot analysis of GSTA and NQO1 in Atg7 +/+ and atg7 −/− macrophages (NS, not significant; n = 2 experiments in duplicate; Student t test). ( C ) Atg7 +/+ and atg7 −/− macrophages were treated with 50 µmol/l tBHP for 24 h (***, P

    Article Snippet: Membranes were probed with the following primary antibodies: goat anti-GSTA (ab53940), rabbit anti-NFE2L2 (ab137550), mouse anti-CDKN2A/p16 (ab54210) and rabbit anti-CDKN1A/p21 (ab7960) from Abcam; rabbit anti-CXCL12 (Bioss, bs-4938); rabbit anti-GAPDH (14C10), rabbit anti-TGFB (3711) and rabbit anti-phospho RB (8516) from Cell Signaling Technology; mouse anti-MAP1LC3B (Nanotools, clone 5F10, 0231-100); rabbit anti-NQO1 (Novus Biologicals, NBP1-40663); rabbit anti-PARP1 (sc-7150), rabbit anti-CDKN2A/p16 (sc-1207) and rabbit anti-total RB (sc-50) from Santa Cruz Biotechnology; mouse anti-ACTB (clone AC-15, A5441), rabbit anti-ATG7 (A2856), rabbit anti-ATG5 (A0856), rabbit anti-SQSTM1/p62 (P0067) and rabbit anti-acetyl-TP53 (SAB4503014) from Sigma-Aldrich.

    Techniques: Derivative Assay, Isolation, Mouse Assay, Western Blot, Quantitative RT-PCR

    Overview of the mechanisms by which defective VSMC autophagy accelerates senescence and promotes postinjury neointima formation and diet-induced atherogenesis. SQSTM1 accumulation in autophagy defective VSMCs triggers NFE2L2 activation and transcription of multiple antioxidative enzymes including GSTA and NQO1. Upregulation of GSTA and NQO1 promotes VSMC survival against oxidative stress under defective autophagy conditions. SQSTM1 accumulation in autophagy defective VSMCs triggers the development of stress-induced premature senescence. Autophagy defective VSMCs are characterized by CDKN2A-RB-mediated G 1 proliferation arrest, increased migration and changes in VSMC phenotype. Enhanced migration is associated with increased secretion of MMP9, TGFB and CXCL12. The phenotype of autophagy defective VSMCs is defined by nuclear and cellular hypertrophy, and by increased collagen content. Defective autophagy in VSMCs accelerates postinjury neointima formation and diet-induced atherogenesis.

    Journal: Autophagy

    Article Title: Defective autophagy in vascular smooth muscle cells accelerates senescence and promotes neointima formation and atherogenesis

    doi: 10.1080/15548627.2015.1096485

    Figure Lengend Snippet: Overview of the mechanisms by which defective VSMC autophagy accelerates senescence and promotes postinjury neointima formation and diet-induced atherogenesis. SQSTM1 accumulation in autophagy defective VSMCs triggers NFE2L2 activation and transcription of multiple antioxidative enzymes including GSTA and NQO1. Upregulation of GSTA and NQO1 promotes VSMC survival against oxidative stress under defective autophagy conditions. SQSTM1 accumulation in autophagy defective VSMCs triggers the development of stress-induced premature senescence. Autophagy defective VSMCs are characterized by CDKN2A-RB-mediated G 1 proliferation arrest, increased migration and changes in VSMC phenotype. Enhanced migration is associated with increased secretion of MMP9, TGFB and CXCL12. The phenotype of autophagy defective VSMCs is defined by nuclear and cellular hypertrophy, and by increased collagen content. Defective autophagy in VSMCs accelerates postinjury neointima formation and diet-induced atherogenesis.

    Article Snippet: Membranes were probed with the following primary antibodies: goat anti-GSTA (ab53940), rabbit anti-NFE2L2 (ab137550), mouse anti-CDKN2A/p16 (ab54210) and rabbit anti-CDKN1A/p21 (ab7960) from Abcam; rabbit anti-CXCL12 (Bioss, bs-4938); rabbit anti-GAPDH (14C10), rabbit anti-TGFB (3711) and rabbit anti-phospho RB (8516) from Cell Signaling Technology; mouse anti-MAP1LC3B (Nanotools, clone 5F10, 0231-100); rabbit anti-NQO1 (Novus Biologicals, NBP1-40663); rabbit anti-PARP1 (sc-7150), rabbit anti-CDKN2A/p16 (sc-1207) and rabbit anti-total RB (sc-50) from Santa Cruz Biotechnology; mouse anti-ACTB (clone AC-15, A5441), rabbit anti-ATG7 (A2856), rabbit anti-ATG5 (A0856), rabbit anti-SQSTM1/p62 (P0067) and rabbit anti-acetyl-TP53 (SAB4503014) from Sigma-Aldrich.

    Techniques: Activation Assay, Migration

    NQO1 protein expression and activity in kidneys from wild-type and Nrf2-null mice 24 h after FeNTA (5 mg/kg) treatment

    Journal:

    Article Title: Coordinated Induction of Nrf2 Target Genes Protects Against Iron Nitrilotriacetate (FeNTA)-Induced Nephrotoxicity

    doi: 10.1016/j.taap.2008.05.022

    Figure Lengend Snippet: NQO1 protein expression and activity in kidneys from wild-type and Nrf2-null mice 24 h after FeNTA (5 mg/kg) treatment

    Article Snippet: Western blots using an anti-NQO1 antibody confirmed that the changes in NQO1 mRNA expression observed in kidneys after FeNTA resulted in elevated protein levels.

    Techniques: Expressing, Activity Assay, Mouse Assay

    Robo-2 is required for the targeting of MOR174–9-expressing OSN axons in the OB. A–J , Coronal sections of OBs isolated from control ( A , D , G , H ) and robo-2 lox/lox ;OMP-Cre; MOR174–9-tau-GFP ( B , C , E , F , I , J ) mice stained with NQO-1 and GFP antibodies ( A–J ). NQO-1 antibodies stain axons of OSNs innervating the dorsal region of the OB. On the lateral side of the OB, a GFP-positive glomerulus is consistently observed in the dorsal region of the OB ( A–C ) and a second GFP-positive glomerulus is observed in approximately half of the OBs analyzed (14/28 controls and 18/25 robo-2 lox/lox ; OMP-Cre ) ( D–F ). In addition, a third glomerulus was observed more ventrally in two OBs from control mice and in six OBs from robo-2 lox/lox ; OMP-Cre mice (shown in yellow on the scatter plot in K ). On the medial side of the OB, all OBs analyzed contained at least one GFP-positive glomerulus ( G , H ) and the majority of OBs analyzed (19/27 for controls and 19/25 for robo-2 lox/lox ; OMP-Cre mice) contained two GFP-positive glomeruli ( H , J ). Furthermore, a few OBs analyzed contained an additional GFP-positive glomerulus (2/28 controls and 6/27 robo-2 lox/lox ; OMP-Cre ). K , L , The relative positions of GFP-positive glomeruli in the dorsoventral axis of the OB on the lateral and medial sides of the OB were assessed and represented on scatter plots ( K , L ). Parameters are shown as mean ± SEM. The mean location of glomeruli innervated by MOR174–9-expressing axons on both the lateral and medial sides of the OB is shifted ventrally in robo-2 lox/lox ; OMP-Cre . Note that the additional glomeruli observed in some bulbs (represented in yellow) were not included in the calculation of the mean. Sections containing glomeruli located close to the mean angle are shown in A , B , D , E , G–J . In C and F , sections with a more severe ventral shift in the location of the glomeruli are shown. *** p

    Journal: The Journal of Neuroscience

    Article Title: The Pattern of Glomerular Map Formation Defines Responsiveness to Aversive Odorants in Mice

    doi: 10.1523/JNEUROSCI.2460-10.2011

    Figure Lengend Snippet: Robo-2 is required for the targeting of MOR174–9-expressing OSN axons in the OB. A–J , Coronal sections of OBs isolated from control ( A , D , G , H ) and robo-2 lox/lox ;OMP-Cre; MOR174–9-tau-GFP ( B , C , E , F , I , J ) mice stained with NQO-1 and GFP antibodies ( A–J ). NQO-1 antibodies stain axons of OSNs innervating the dorsal region of the OB. On the lateral side of the OB, a GFP-positive glomerulus is consistently observed in the dorsal region of the OB ( A–C ) and a second GFP-positive glomerulus is observed in approximately half of the OBs analyzed (14/28 controls and 18/25 robo-2 lox/lox ; OMP-Cre ) ( D–F ). In addition, a third glomerulus was observed more ventrally in two OBs from control mice and in six OBs from robo-2 lox/lox ; OMP-Cre mice (shown in yellow on the scatter plot in K ). On the medial side of the OB, all OBs analyzed contained at least one GFP-positive glomerulus ( G , H ) and the majority of OBs analyzed (19/27 for controls and 19/25 for robo-2 lox/lox ; OMP-Cre mice) contained two GFP-positive glomeruli ( H , J ). Furthermore, a few OBs analyzed contained an additional GFP-positive glomerulus (2/28 controls and 6/27 robo-2 lox/lox ; OMP-Cre ). K , L , The relative positions of GFP-positive glomeruli in the dorsoventral axis of the OB on the lateral and medial sides of the OB were assessed and represented on scatter plots ( K , L ). Parameters are shown as mean ± SEM. The mean location of glomeruli innervated by MOR174–9-expressing axons on both the lateral and medial sides of the OB is shifted ventrally in robo-2 lox/lox ; OMP-Cre . Note that the additional glomeruli observed in some bulbs (represented in yellow) were not included in the calculation of the mean. Sections containing glomeruli located close to the mean angle are shown in A , B , D , E , G–J . In C and F , sections with a more severe ventral shift in the location of the glomeruli are shown. *** p

    Article Snippet: Sections were immunostained with NQO-1 antibodies (1:100; Abcam), and GFP (1:250; Invitrogen) or β-gal (1:250; MP Biomedicals) antibodies, and counterstained with Hoechst.

    Techniques: Expressing, Isolation, Mouse Assay, Staining

    MOR1–3-expressing OSN axons coalesce accurately in robo-2 lox/lox ; OMP-Cre mice. A–D , Coronal sections of OBs isolated from control ( A , C ) and robo-2 lox/lox ;OMP-Cre; MOR1–3-tau-lacZ ( B , D ) mice stained with NQO-1 and β-galactosidase antibodies ( A–D ). A β-galactosidase-positive glomerulus is consistently observed in the dorsomedial part of the OB in all mice analyzed ( C , D ). In addition, a second β-galactosidase-positive glomerulus is observed on the dorsal part of the OB in approximately half of the OBs analyzed (9/17 controls and 12/20 robo-2 lox/lox ; OMP-Cre ) ( A , B ). E , The relative positions of β-galactosidase-positive glomeruli in the dorsoventral axis of the OB was assessed and represented on a scatter plot. Parameters are shown as mean ± SEM. No significant change was observed in the mean location of MOR1–3-positive glomeruli in robo-2 lox/lox ; OMP-Cre mice. Note that in A–D sections with glomeruli located approximately at the mean angle are shown. D, Dorsal; V, ventral; L, lateral; M, medial. Scale bar, 500 μm.

    Journal: The Journal of Neuroscience

    Article Title: The Pattern of Glomerular Map Formation Defines Responsiveness to Aversive Odorants in Mice

    doi: 10.1523/JNEUROSCI.2460-10.2011

    Figure Lengend Snippet: MOR1–3-expressing OSN axons coalesce accurately in robo-2 lox/lox ; OMP-Cre mice. A–D , Coronal sections of OBs isolated from control ( A , C ) and robo-2 lox/lox ;OMP-Cre; MOR1–3-tau-lacZ ( B , D ) mice stained with NQO-1 and β-galactosidase antibodies ( A–D ). A β-galactosidase-positive glomerulus is consistently observed in the dorsomedial part of the OB in all mice analyzed ( C , D ). In addition, a second β-galactosidase-positive glomerulus is observed on the dorsal part of the OB in approximately half of the OBs analyzed (9/17 controls and 12/20 robo-2 lox/lox ; OMP-Cre ) ( A , B ). E , The relative positions of β-galactosidase-positive glomeruli in the dorsoventral axis of the OB was assessed and represented on a scatter plot. Parameters are shown as mean ± SEM. No significant change was observed in the mean location of MOR1–3-positive glomeruli in robo-2 lox/lox ; OMP-Cre mice. Note that in A–D sections with glomeruli located approximately at the mean angle are shown. D, Dorsal; V, ventral; L, lateral; M, medial. Scale bar, 500 μm.

    Article Snippet: Sections were immunostained with NQO-1 antibodies (1:100; Abcam), and GFP (1:250; Invitrogen) or β-gal (1:250; MP Biomedicals) antibodies, and counterstained with Hoechst.

    Techniques: Expressing, Mouse Assay, Isolation, Staining

    Expression of Robo family members in the olfactory epithelium. In situ hybridization of coronal sections of olfactory epithelium isolated from E16 embryos ( A–E ) and adult mice (P90) ( F–M ) with cRNA probes specific for robo-1 ( A ), rig-1 ( B ), robo-2 ( C , G , H ), NQO-1 ( D , I ), OCAM ( E , J ), OMP ( F ), M49 ( K ), L45 ( L ), and M50 ( M ). A–G , Expression of robo-1 is restricted to the basal lamina of the olfactory epithelium ( A , arrowheads), whereas rig-1 is not expressed in the olfactory epithelium ( B ). Interestingly, in contrast to OMP, which is equally expressed throughout the olfactory epithelium, robo-2 is expressed in a gradient with high levels of expression in the dorsomedial regions of the olfactory epithelium (arrows) and low levels in the ventrolateral regions (arrowheads) of E16 ( C ) and adult olfactory epithelium ( G , H ). H–M , robo-2 expression is confined to zones I–III of the olfactory epithelium. A higher magnification of a region of the olfactory epithelium (boxed in G ) in which all four zones of the olfactory epithelium are represented is shown in H–M . The expression pattern of robo-2 was compared with the expression pattern of specific olfactory epithelium zonal markers that include NQO-1 (zone I) ( I ), OCAM (zones II–IV) ( J ), M49 (zone II) ( K ), L45 (zone III) ( L ), and M50 (zone IV) ( M ). robo-2 is expressed in a high-to-low gradient in olfactory sensory neurons located in zones I (arrows) to IV (arrowheads), respectively. Regions of the olfactory epithelium expressing the different zonal markers are traced with a colored line on the apical surface of the olfactory epithelium to represent the four zones (zone I, magenta; zone II, green; zone III, yellow; zone IV, blue). S, Septum. Scale bars: A–E , H–M , 250 μm; F , G , 500 μm. N , Diagram representing the spatial relationship between the location of olfactory sensory neurons within the OE and their target glomeruli within the OB. Olfactory sensory neurons located in the dorsomedial regions of the olfactory epithelium (magenta) project axons to glomeruli in the dorsal region of the olfactory bulb, whereas olfactory sensory neurons in the ventrolateral region of the olfactory epithelium (blue and not shown) project axons to the ventral aspect of the olfactory bulb. D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Expression of Robo family members in the olfactory epithelium. In situ hybridization of coronal sections of olfactory epithelium isolated from E16 embryos ( A–E ) and adult mice (P90) ( F–M ) with cRNA probes specific for robo-1 ( A ), rig-1 ( B ), robo-2 ( C , G , H ), NQO-1 ( D , I ), OCAM ( E , J ), OMP ( F ), M49 ( K ), L45 ( L ), and M50 ( M ). A–G , Expression of robo-1 is restricted to the basal lamina of the olfactory epithelium ( A , arrowheads), whereas rig-1 is not expressed in the olfactory epithelium ( B ). Interestingly, in contrast to OMP, which is equally expressed throughout the olfactory epithelium, robo-2 is expressed in a gradient with high levels of expression in the dorsomedial regions of the olfactory epithelium (arrows) and low levels in the ventrolateral regions (arrowheads) of E16 ( C ) and adult olfactory epithelium ( G , H ). H–M , robo-2 expression is confined to zones I–III of the olfactory epithelium. A higher magnification of a region of the olfactory epithelium (boxed in G ) in which all four zones of the olfactory epithelium are represented is shown in H–M . The expression pattern of robo-2 was compared with the expression pattern of specific olfactory epithelium zonal markers that include NQO-1 (zone I) ( I ), OCAM (zones II–IV) ( J ), M49 (zone II) ( K ), L45 (zone III) ( L ), and M50 (zone IV) ( M ). robo-2 is expressed in a high-to-low gradient in olfactory sensory neurons located in zones I (arrows) to IV (arrowheads), respectively. Regions of the olfactory epithelium expressing the different zonal markers are traced with a colored line on the apical surface of the olfactory epithelium to represent the four zones (zone I, magenta; zone II, green; zone III, yellow; zone IV, blue). S, Septum. Scale bars: A–E , H–M , 250 μm; F , G , 500 μm. N , Diagram representing the spatial relationship between the location of olfactory sensory neurons within the OE and their target glomeruli within the OB. Olfactory sensory neurons located in the dorsomedial regions of the olfactory epithelium (magenta) project axons to glomeruli in the dorsal region of the olfactory bulb, whereas olfactory sensory neurons in the ventrolateral region of the olfactory epithelium (blue and not shown) project axons to the ventral aspect of the olfactory bulb. D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Expressing, In Situ Hybridization, Isolation, Mouse Assay

    Olfactory sensory neuron projections are disorganized in robo-2 −/− mice. Parasagittal ( A–F ) and coronal ( G–O ) sections of olfactory bulbs from P0 robo-2 +/+ ( A–C , G–I ) and robo-2 −/− ( D–F , J–O ) mice were stained with anti-NQO-1 ( A , C , D , F , G , I , J , L , M , O ) and anti-OCAM ( B , C , E , F , H , I , K , L , N , O ). In robo-2 −/− mice, NQO-1-expressing axons are restricted to the rostral and dorsal region of the olfactory bulbs ( A , G ), whereas OCAM-expressing axons target the ventral region of the olfactory bulbs ( B , H ). A subset of NQO-1-expressing axons is mistargeted to the ventral region of the olfactory bulbs in robo-2 −/− mice (arrowheads) ( D , F , J , L , M , O ). In addition, some regions of the ventral olfactory bulb that are innervated by OCAM-expressing axons in robo-2 +/+ mice lack innervation in robo-2 −/− mice (arrows). n = 9 robo-2 +/+ , 3 robo-2 +/− , and 11 robo-2 −/− . D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal. Scale bars, 250 μm.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Olfactory sensory neuron projections are disorganized in robo-2 −/− mice. Parasagittal ( A–F ) and coronal ( G–O ) sections of olfactory bulbs from P0 robo-2 +/+ ( A–C , G–I ) and robo-2 −/− ( D–F , J–O ) mice were stained with anti-NQO-1 ( A , C , D , F , G , I , J , L , M , O ) and anti-OCAM ( B , C , E , F , H , I , K , L , N , O ). In robo-2 −/− mice, NQO-1-expressing axons are restricted to the rostral and dorsal region of the olfactory bulbs ( A , G ), whereas OCAM-expressing axons target the ventral region of the olfactory bulbs ( B , H ). A subset of NQO-1-expressing axons is mistargeted to the ventral region of the olfactory bulbs in robo-2 −/− mice (arrowheads) ( D , F , J , L , M , O ). In addition, some regions of the ventral olfactory bulb that are innervated by OCAM-expressing axons in robo-2 +/+ mice lack innervation in robo-2 −/− mice (arrows). n = 9 robo-2 +/+ , 3 robo-2 +/− , and 11 robo-2 −/− . D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal. Scale bars, 250 μm.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Mouse Assay, Staining, Expressing

    Expression of NQO-1 and OCAM are unaltered in robo-2 −/− and slit-1 −/− ; slit-3 −/− mice. A–N , In situ hybridization of coronal sections of olfactory epithelia from P0 wild-type ( A , D , G , I , L ), slit-1 −/− ; slit-3 −/− ( B , E , H , J , M ), and robo-2 −/− ( C , F , K , N ) mice with cRNA probes specific for NQO-1 ( A–C ), OCAM ( D–F ), robo-2 ( G , H ), robo-1 ( I–K ), and rig-1 ( L–N ). robo-2 is expressed in a gradient in the OE with high levels of expression in the dorsomedial region of the OE and low levels in the ventrolateral region in wild-type ( G ) and slit-1 −/− ; slit-3 −/− ( H ) mice. NQO-1 is expressed in the dorsomedial region, whereas OCAM is expressed in the ventrolateral region of the olfactory epithelium in wild-type ( A , D ), slit-1 −/− ; slit-3 −/− ( B , E ), and robo-2 −/− ( C , F ) mice. Both robo-1 and rig-1 are not expressed in the olfactory epithelium in wild-type ( I , L ), slit-1 −/− ; slit-3 −/− ( J , M ), and robo-2 −/− ( K , N ) mice. n = 5 robo-2 −/− and n = 5 slit-1 −/− ; slit-3 −/− . O , P , In situ hybridization of parasagittal sections of olfactory bulbs from E16 slit-1 −/− embryos with cRNA probes specific for slit-3 ( O ) and slit-2 ( P ). Whereas slit-3 is expressed in the ventral region of the olfactory bulb, slit-2 is not expressed in the olfactory bulb of slit-1 −/− embryos. n = 3 slit-1 −/− . D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal. Scale bars, 250 μm.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Expression of NQO-1 and OCAM are unaltered in robo-2 −/− and slit-1 −/− ; slit-3 −/− mice. A–N , In situ hybridization of coronal sections of olfactory epithelia from P0 wild-type ( A , D , G , I , L ), slit-1 −/− ; slit-3 −/− ( B , E , H , J , M ), and robo-2 −/− ( C , F , K , N ) mice with cRNA probes specific for NQO-1 ( A–C ), OCAM ( D–F ), robo-2 ( G , H ), robo-1 ( I–K ), and rig-1 ( L–N ). robo-2 is expressed in a gradient in the OE with high levels of expression in the dorsomedial region of the OE and low levels in the ventrolateral region in wild-type ( G ) and slit-1 −/− ; slit-3 −/− ( H ) mice. NQO-1 is expressed in the dorsomedial region, whereas OCAM is expressed in the ventrolateral region of the olfactory epithelium in wild-type ( A , D ), slit-1 −/− ; slit-3 −/− ( B , E ), and robo-2 −/− ( C , F ) mice. Both robo-1 and rig-1 are not expressed in the olfactory epithelium in wild-type ( I , L ), slit-1 −/− ; slit-3 −/− ( J , M ), and robo-2 −/− ( K , N ) mice. n = 5 robo-2 −/− and n = 5 slit-1 −/− ; slit-3 −/− . O , P , In situ hybridization of parasagittal sections of olfactory bulbs from E16 slit-1 −/− embryos with cRNA probes specific for slit-3 ( O ) and slit-2 ( P ). Whereas slit-3 is expressed in the ventral region of the olfactory bulb, slit-2 is not expressed in the olfactory bulb of slit-1 −/− embryos. n = 3 slit-1 −/− . D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal. Scale bars, 250 μm.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Expressing, Mouse Assay, In Situ Hybridization

    Targeting of Robo-2-expressing olfactory sensory neuron axons in the olfactory bulb. A–L , Parasagittal sections of olfactory bulbs from E16 ( A–F ) and E18 ( G–L ) embryos stained with anti-Robo-2 ( A , C , D , F , G , I , J , L ), anti-NQO-1 ( B , C , H , I ), and anti-OCAM ( E , F , K , L ). At E16 and E18, olfactory sensory neuron axons expressing high levels of Robo-2 are observed in the dorsal and rostral regions of the OB ( A , D , G , J ). Robo-2 is also expressed at lower levels on a subset of axons targeting the ventral region of the olfactory bulb. NQO-1-expressing axons, originating from zone I of the olfactory epithelium, that target to the dorsal region of the OB express high levels of Robo-2 ( B , C , H , I ) (arrowheads). A subset of OCAM-expressing axons that originate from zones II–IV of the olfactory epithelium and target to the ventral region of the olfactory bulb do not express Robo-2 (arrow) ( E , F , K , L ). M–R , Coronal sections of olfactory bulbs from E18 embryos stained with anti-Robo-2 ( M , O , P , R ), anti-NQO-1 ( N , O ), and anti-OCAM ( Q , R ). At E18, olfactory sensory neuron axons expressing high levels of Robo-2 are observed in the dorsomedial regions of the OB (arrowheads), whereas lower levels of Robo-2 expression are observed on olfactory sensory neuron axons targeting to the ventrolateral region of the OB. As observed in sagittal sections ( G–L ), NQO-1-positive axons express high levels of Robo-2 (arrowheads) ( M–O ), whereas a subset of OCAM-positive axons do not express Robo-2 (arrows) ( P–R ). Robo-2-positive axons restricted to the nerve layer are marked with asterisks in P and R . D, Dorsal; V, ventral; R, rostral; C, caudal; M, medial; L, lateral. Scale bars, 250 μm.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Targeting of Robo-2-expressing olfactory sensory neuron axons in the olfactory bulb. A–L , Parasagittal sections of olfactory bulbs from E16 ( A–F ) and E18 ( G–L ) embryos stained with anti-Robo-2 ( A , C , D , F , G , I , J , L ), anti-NQO-1 ( B , C , H , I ), and anti-OCAM ( E , F , K , L ). At E16 and E18, olfactory sensory neuron axons expressing high levels of Robo-2 are observed in the dorsal and rostral regions of the OB ( A , D , G , J ). Robo-2 is also expressed at lower levels on a subset of axons targeting the ventral region of the olfactory bulb. NQO-1-expressing axons, originating from zone I of the olfactory epithelium, that target to the dorsal region of the OB express high levels of Robo-2 ( B , C , H , I ) (arrowheads). A subset of OCAM-expressing axons that originate from zones II–IV of the olfactory epithelium and target to the ventral region of the olfactory bulb do not express Robo-2 (arrow) ( E , F , K , L ). M–R , Coronal sections of olfactory bulbs from E18 embryos stained with anti-Robo-2 ( M , O , P , R ), anti-NQO-1 ( N , O ), and anti-OCAM ( Q , R ). At E18, olfactory sensory neuron axons expressing high levels of Robo-2 are observed in the dorsomedial regions of the OB (arrowheads), whereas lower levels of Robo-2 expression are observed on olfactory sensory neuron axons targeting to the ventrolateral region of the OB. As observed in sagittal sections ( G–L ), NQO-1-positive axons express high levels of Robo-2 (arrowheads) ( M–O ), whereas a subset of OCAM-positive axons do not express Robo-2 (arrows) ( P–R ). Robo-2-positive axons restricted to the nerve layer are marked with asterisks in P and R . D, Dorsal; V, ventral; R, rostral; C, caudal; M, medial; L, lateral. Scale bars, 250 μm.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Expressing, Staining

    Mistargeted axons of NQO-1-expressing olfactory sensory neurons innervate glomeruli in the ventral region of the olfactory bulb in slit and robo-2 mutant mice. Coronal sections at similar rostrocaudal levels of olfactory bulbs isolated from 8- to 12-week-old wild-type ( A , E , I ), slit-1 −/− ( B , F , J , M–O ), slit-1 −/− ; slit-3 −/− ( C , G , K ), and robo-2 c/c ; syn-1 Cre /+ ( D , H , L ) mice stained with anti-NQO-1 ( A–D , I–L , M–O ), anti-OCAM ( E–H , I–L , M , N ), anti-SV2 ( O ), and Hoechst ( A–O ). In wild-type mice, NQO-1-expressing axons innervate glomeruli restricted to the dorsomedial region of the olfactory bulb ( A , I ), and OCAM-expressing axons innervate glomeruli in the ventrolateral region of the olfactory bulb ( E , I ). In slit-1 −/− ( B , J ), slit-1 −/− ; slit-3 −/− ( C , K ), and robo-2 c/c ;syn-1 Cre /+ ( D , L ) mice, a subset of NQO-1-expressing axons are mistargeted to glomeruli in the ventral region of the olfactory bulb (arrows). NQO-1-expressing axons that target ectopically in the ventral region of the OB in slit-1 −/− mice form both homogenous (arrowheads) and heterogenous (arrows) glomeruli ( M , N ) and are positive for the presynaptic marker SV2, suggesting that they form synapses (arrowheads) ( O ). n = 10 wild type, n = 14 slit-1 −/− , n = 9 slit-1 −/− ; slit-3 −/− , and n = 4 robo-2 c/c ;syn-1 Cre /+ . Scale bars: A–L , 250 μm; M–O , 140 μm. P–R , Scatter plots showing the mapping of the positions of NQO-1-positive glomeruli in the olfactory bulb of an adult wild-type ( P ), slit-1 −/− ( Q ), and robo-2 c/c ;syn-1 Cre /+ ( R ) mouse. The relative positions of glomeruli containing NQO-1-positive axons were assessed in olfactory bulb sections isolated over a rostrocaudal distance of 1000 μm starting at 800 μm from the tip of the olfactory bulb. Although NQO-1-positive glomeruli are absent in the ventral region of the olfactory bulb (180° angle) from a wild-type mouse, NQO-1-positive glomeruli are consistently observed in the most ventral region of the OB in slit-1 −/− and robo-2 c/c ;syn-1 Cre /+ mice. Shown are representative plots from a single olfactory bulb for each genotype ( n = 4 olfactory bulbs from 2 mice of each genotype). D, Dorsal; V, ventral; L, lateral; M, medial.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Mistargeted axons of NQO-1-expressing olfactory sensory neurons innervate glomeruli in the ventral region of the olfactory bulb in slit and robo-2 mutant mice. Coronal sections at similar rostrocaudal levels of olfactory bulbs isolated from 8- to 12-week-old wild-type ( A , E , I ), slit-1 −/− ( B , F , J , M–O ), slit-1 −/− ; slit-3 −/− ( C , G , K ), and robo-2 c/c ; syn-1 Cre /+ ( D , H , L ) mice stained with anti-NQO-1 ( A–D , I–L , M–O ), anti-OCAM ( E–H , I–L , M , N ), anti-SV2 ( O ), and Hoechst ( A–O ). In wild-type mice, NQO-1-expressing axons innervate glomeruli restricted to the dorsomedial region of the olfactory bulb ( A , I ), and OCAM-expressing axons innervate glomeruli in the ventrolateral region of the olfactory bulb ( E , I ). In slit-1 −/− ( B , J ), slit-1 −/− ; slit-3 −/− ( C , K ), and robo-2 c/c ;syn-1 Cre /+ ( D , L ) mice, a subset of NQO-1-expressing axons are mistargeted to glomeruli in the ventral region of the olfactory bulb (arrows). NQO-1-expressing axons that target ectopically in the ventral region of the OB in slit-1 −/− mice form both homogenous (arrowheads) and heterogenous (arrows) glomeruli ( M , N ) and are positive for the presynaptic marker SV2, suggesting that they form synapses (arrowheads) ( O ). n = 10 wild type, n = 14 slit-1 −/− , n = 9 slit-1 −/− ; slit-3 −/− , and n = 4 robo-2 c/c ;syn-1 Cre /+ . Scale bars: A–L , 250 μm; M–O , 140 μm. P–R , Scatter plots showing the mapping of the positions of NQO-1-positive glomeruli in the olfactory bulb of an adult wild-type ( P ), slit-1 −/− ( Q ), and robo-2 c/c ;syn-1 Cre /+ ( R ) mouse. The relative positions of glomeruli containing NQO-1-positive axons were assessed in olfactory bulb sections isolated over a rostrocaudal distance of 1000 μm starting at 800 μm from the tip of the olfactory bulb. Although NQO-1-positive glomeruli are absent in the ventral region of the olfactory bulb (180° angle) from a wild-type mouse, NQO-1-positive glomeruli are consistently observed in the most ventral region of the OB in slit-1 −/− and robo-2 c/c ;syn-1 Cre /+ mice. Shown are representative plots from a single olfactory bulb for each genotype ( n = 4 olfactory bulbs from 2 mice of each genotype). D, Dorsal; V, ventral; L, lateral; M, medial.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Expressing, Mutagenesis, Mouse Assay, Isolation, Staining, Marker

    Slit-1 and Robo-2 are required for the segregation of zone I OSN axons to the dorsal region of the OB. Representation of zone I OSN projections in wild-type, slit-1 −/− , and robo-2 −/− mice. NQO-1-expressing zone I OSNs (magenta) located in the dorsal region of the OE project their axons to the dorsal aspect of the OB. The high dorsal to low ventral graded expression of Robo-2 (green) in OSN of the OE promotes the segregation of NQO-1-expressing axons to the dorsal region of the OB. Zone I OSN axons may be repelled by the Robo-2 ligand Slit-1, which is expressed in the ventral region of the OB. Loss of either Robo-2 or Slit-1 expression leads to mistargeting of a subset of zone I OSN axons to the ventral aspect of the OB.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Slit-1 and Robo-2 are required for the segregation of zone I OSN axons to the dorsal region of the OB. Representation of zone I OSN projections in wild-type, slit-1 −/− , and robo-2 −/− mice. NQO-1-expressing zone I OSNs (magenta) located in the dorsal region of the OE project their axons to the dorsal aspect of the OB. The high dorsal to low ventral graded expression of Robo-2 (green) in OSN of the OE promotes the segregation of NQO-1-expressing axons to the dorsal region of the OB. Zone I OSN axons may be repelled by the Robo-2 ligand Slit-1, which is expressed in the ventral region of the OB. Loss of either Robo-2 or Slit-1 expression leads to mistargeting of a subset of zone I OSN axons to the ventral aspect of the OB.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Mouse Assay, Expressing

    Loss of zonal targeting of NQO-1-expressing olfactory sensory neuron axons within the olfactory bulb in slit-1 −/− mice. A–M , Parasagittal sections of olfactory bulbs from P0 wild-type ( A–C ), slit-1 −/− ( D–F ), slit-3 −/− ( H–J ), and slit-1 −/− ; slit-3 −/− ( K–M ) mice stained with anti-NQO-1 ( A , C , D , F , H , J , K , M ) and anti-OCAM ( B , C , E , F , I , J , L , M ). In wild-type animals, NQO-1-expressing axons are restricted to the rostrodorsal region of the OB ( A , C ) and OCAM-expressing axons target to the ventral region of the olfactory bulb ( B , C ). Whereas NQO-1-expressing axons are properly targeted to the dorsal region of the olfactory bulb in slit-3 −/− mice ( H , J ), a subset of NQO-1-expressing axons mistarget to the most ventral region of the olfactory bulb in slit-1 −/− mice ( D , F ) (arrowheads). In slit-1 −/− ; slit-3 −/− mice, NQO-1-expressing axons are also observed in the ventral region of the olfactory bulb ( K , M ) (arrowheads). High-powered magnifications of ectopically projecting NQO-1-expressing axons (insets in D , F , K , M ) are shown in D′ , F′ , K′ , and M′ . n = 8 wild type, n = 5 slit-1 −/− , n = 8 slit-3 −/− , and n = 7 slit-1 −/− ; slit-3 −/− . N–P , Coronal sections at similar rostrocaudal levels of olfactory bulbs isolated from P0 ( N–P ) wild-type ( N ), slit-1 −/− ( O ), and slit-1 −/− ; slit-3 −/− ( P ) mice stained with anti-NQO-1 ( N–P ). In wild-type mice, NQO-1-expressing axons innervate the dorsomedial region of the olfactory bulb. In slit-1 −/− and slit-1 −/− ; slit-3 −/− mice, a subset of NQO-1-expressing axons are mistargeted to the ventral region of the olfactory bulb (arrows). n = 8 wild type, n = 8 slit-1 −/− , and n = 5 slit-1 −/− ; slit-3 −/− . D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal. Scale bars, 250 μm.

    Journal: The Journal of Neuroscience

    Article Title: Requirement for Slit-1 and Robo-2 in Zonal Segregation of Olfactory Sensory Neuron Axons in the Main Olfactory Bulb

    doi: 10.1523/JNEUROSCI.2217-07.2007

    Figure Lengend Snippet: Loss of zonal targeting of NQO-1-expressing olfactory sensory neuron axons within the olfactory bulb in slit-1 −/− mice. A–M , Parasagittal sections of olfactory bulbs from P0 wild-type ( A–C ), slit-1 −/− ( D–F ), slit-3 −/− ( H–J ), and slit-1 −/− ; slit-3 −/− ( K–M ) mice stained with anti-NQO-1 ( A , C , D , F , H , J , K , M ) and anti-OCAM ( B , C , E , F , I , J , L , M ). In wild-type animals, NQO-1-expressing axons are restricted to the rostrodorsal region of the OB ( A , C ) and OCAM-expressing axons target to the ventral region of the olfactory bulb ( B , C ). Whereas NQO-1-expressing axons are properly targeted to the dorsal region of the olfactory bulb in slit-3 −/− mice ( H , J ), a subset of NQO-1-expressing axons mistarget to the most ventral region of the olfactory bulb in slit-1 −/− mice ( D , F ) (arrowheads). In slit-1 −/− ; slit-3 −/− mice, NQO-1-expressing axons are also observed in the ventral region of the olfactory bulb ( K , M ) (arrowheads). High-powered magnifications of ectopically projecting NQO-1-expressing axons (insets in D , F , K , M ) are shown in D′ , F′ , K′ , and M′ . n = 8 wild type, n = 5 slit-1 −/− , n = 8 slit-3 −/− , and n = 7 slit-1 −/− ; slit-3 −/− . N–P , Coronal sections at similar rostrocaudal levels of olfactory bulbs isolated from P0 ( N–P ) wild-type ( N ), slit-1 −/− ( O ), and slit-1 −/− ; slit-3 −/− ( P ) mice stained with anti-NQO-1 ( N–P ). In wild-type mice, NQO-1-expressing axons innervate the dorsomedial region of the olfactory bulb. In slit-1 −/− and slit-1 −/− ; slit-3 −/− mice, a subset of NQO-1-expressing axons are mistargeted to the ventral region of the olfactory bulb (arrows). n = 8 wild type, n = 8 slit-1 −/− , and n = 5 slit-1 −/− ; slit-3 −/− . D, Dorsal; V, ventral; L, lateral; M, medial; R, rostral; C, caudal. Scale bars, 250 μm.

    Article Snippet: The sections were then blocked for 2 h in TNT (50 m m Tris-HCl, pH 7.6, 500 m m NaCl, and 0.5% Triton X-100) containing 10% fetal bovine serum (FBS) and incubated overnight with primary antibody at 4°C in TNT/10% FBS using the following dilutions: anti-OCAM at 1:100 (BD Biosciences, San Jose, CA), anti-NQO-1 at 1:100 (Abcam, Cambridge, MA), anti-SV2 (synaptic vesicle protein 2) at 1:1000 (Developmental Studies Hybridoma Bank, University of Iowa, Iowa City, IA), and anti-Robo-2 at 1:350.

    Techniques: Expressing, Mouse Assay, Staining, Isolation

    Nrf2-activators dose-dependently increase antioxidant protein expression in OLN-93 cells. HO-1 ( A ), NQO-1 ( B ) and p62 ( C ) protein expression levels after 24 h treatment in the OLN-93 oligodendrocyte cell line with 5 µM or 10 µM SFN, 45 µM or 90 µM MMF, 30 µg/mL or 60 µg/mL Protandim or their respective DMSO or EtOH vehicle control. Protein levels were assayed by Western blotting. Data are presented as percentage of control and expressed as the mean ± SEM of 3 independent experiments. All statistics reflect one-way ANOVA tests with post hoc Bonferroni correction; * p

    Journal: Antioxidants

    Article Title: Protandim Protects Oligodendrocytes against an Oxidative Insult

    doi: 10.3390/antiox5030030

    Figure Lengend Snippet: Nrf2-activators dose-dependently increase antioxidant protein expression in OLN-93 cells. HO-1 ( A ), NQO-1 ( B ) and p62 ( C ) protein expression levels after 24 h treatment in the OLN-93 oligodendrocyte cell line with 5 µM or 10 µM SFN, 45 µM or 90 µM MMF, 30 µg/mL or 60 µg/mL Protandim or their respective DMSO or EtOH vehicle control. Protein levels were assayed by Western blotting. Data are presented as percentage of control and expressed as the mean ± SEM of 3 independent experiments. All statistics reflect one-way ANOVA tests with post hoc Bonferroni correction; * p

    Article Snippet: Antibody Characterization Primary antibodies used were rat anti-MBP (catalog no. MCA409S; RRID: AB_325004; Abd Serotec, Oxfordshire, UK), rabbit anti-Olig2 (catalog no. AB9610; RRID: AB_570666; Millipore, Billerica, MA, USA), rabbit anti-HO-1 (catalog no. ADI-OSA-150F; RRID: AB_1505620; Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-NQO-1 (catalog no. ab28947; RRID: AB_881738; Abcam, Cambridge, UK), and mouse anti-p62/SQSTM1 (catalog no. ab56416; RRID: AB_945626), mouse anti-β-actin (catalog no. A5441; RRID: AB_476744; Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Expressing, Western Blot

    Nrf2-activators dose-dependently increase antioxidant protein expression in mature primary rat OLs. HO-1, NQO-1 and p62 protein expression levels after 24 h treatment in mature primary rat OLs, differentiated for 7 days, with 5 µM SFN, 90 µM MMF,30 µg/mL Protandim or their respective DMSO or EtOH vehicle control. Protein levels were assayed by Western blotting. Data are presented as percentage of control and expressed as the mean ± SEM of 3 independent experiments. Statistics reflect one-way ANOVA test with post hoc Bonferroni correction; * p

    Journal: Antioxidants

    Article Title: Protandim Protects Oligodendrocytes against an Oxidative Insult

    doi: 10.3390/antiox5030030

    Figure Lengend Snippet: Nrf2-activators dose-dependently increase antioxidant protein expression in mature primary rat OLs. HO-1, NQO-1 and p62 protein expression levels after 24 h treatment in mature primary rat OLs, differentiated for 7 days, with 5 µM SFN, 90 µM MMF,30 µg/mL Protandim or their respective DMSO or EtOH vehicle control. Protein levels were assayed by Western blotting. Data are presented as percentage of control and expressed as the mean ± SEM of 3 independent experiments. Statistics reflect one-way ANOVA test with post hoc Bonferroni correction; * p

    Article Snippet: Antibody Characterization Primary antibodies used were rat anti-MBP (catalog no. MCA409S; RRID: AB_325004; Abd Serotec, Oxfordshire, UK), rabbit anti-Olig2 (catalog no. AB9610; RRID: AB_570666; Millipore, Billerica, MA, USA), rabbit anti-HO-1 (catalog no. ADI-OSA-150F; RRID: AB_1505620; Enzo Life Sciences, Farmingdale, NY, USA), mouse anti-NQO-1 (catalog no. ab28947; RRID: AB_881738; Abcam, Cambridge, UK), and mouse anti-p62/SQSTM1 (catalog no. ab56416; RRID: AB_945626), mouse anti-β-actin (catalog no. A5441; RRID: AB_476744; Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Expressing, Western Blot

    (A) mRNA expression of hepatic Nrf2, NQO-1, ER α , and TFF3. Total RNA was extracted from each liver collected from the experiment illustrated in . Hepatic mRNA expression of the genes indicated was quantified with quantitative real-time PCR.

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title:

    doi: 10.1124/jpet.115.231316

    Figure Lengend Snippet: (A) mRNA expression of hepatic Nrf2, NQO-1, ER α , and TFF3. Total RNA was extracted from each liver collected from the experiment illustrated in . Hepatic mRNA expression of the genes indicated was quantified with quantitative real-time PCR.

    Article Snippet: Antibodies against NQO-1 (ab80588; Abcam), ER α (SC-514910; Santa Cruz Biotechnology, Santa Cruz, CA), FABP5 (GTX12109; GeneTex), and glyceraldehyde-3-phosphate dehydrogenase (SC-130656; Santa Cruz Biotechnology) were used as probes.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Equol attenuated ER stress by activating Nrf2 in vitro and in vivo. (A) HUVECs were incubated with different concentrations (1, 10 and 100 nM) of equol for 24 h before treatment with t -BHP (50 μM) for another 6 h. The expression of Nrf2 and NQO1 was detected by western blotting. (C) Total tissue lysates from the thoracic and abdominal aorta of apoE-/- mice with or without equol treatment were immunoblotted with anti-Nrf2 and anti-β-actin antibodies. (E) Cells were transfected with Nrf2 siRNA for 5~6 h; the medium was then replaced with fresh culture medium, followed by incubation for another 24 h. Thereafter, the cells were treated with equol (100 nM) for 24 h and then were incubated with t -BHP (50 μM) for an additional 6 h. The cells were collected and lysed, and western blot analysis was performed. (B)(D)(F) The bar charts show the quantification of the indicated proteins. (G) HUVECs were transfected with Nrf2 siRNA and treated as described in (C), and cell apoptosis was assayed using the Cell Death Detection ELISA plus Kit. Values are presented as means ± SD. n = 3, * p

    Journal: PLoS ONE

    Article Title: Equol Attenuates Atherosclerosis in Apolipoprotein E-Deficient Mice by Inhibiting Endoplasmic Reticulum Stress via Activation of Nrf2 in Endothelial Cells

    doi: 10.1371/journal.pone.0167020

    Figure Lengend Snippet: Equol attenuated ER stress by activating Nrf2 in vitro and in vivo. (A) HUVECs were incubated with different concentrations (1, 10 and 100 nM) of equol for 24 h before treatment with t -BHP (50 μM) for another 6 h. The expression of Nrf2 and NQO1 was detected by western blotting. (C) Total tissue lysates from the thoracic and abdominal aorta of apoE-/- mice with or without equol treatment were immunoblotted with anti-Nrf2 and anti-β-actin antibodies. (E) Cells were transfected with Nrf2 siRNA for 5~6 h; the medium was then replaced with fresh culture medium, followed by incubation for another 24 h. Thereafter, the cells were treated with equol (100 nM) for 24 h and then were incubated with t -BHP (50 μM) for an additional 6 h. The cells were collected and lysed, and western blot analysis was performed. (B)(D)(F) The bar charts show the quantification of the indicated proteins. (G) HUVECs were transfected with Nrf2 siRNA and treated as described in (C), and cell apoptosis was assayed using the Cell Death Detection ELISA plus Kit. Values are presented as means ± SD. n = 3, * p

    Article Snippet: NQO1 antibody (A180) was purchased from NOVUS Biologicals (Littleton, CO), activated-caspase-3 p17 (BS7004) was obtained from Bioworld Technology, whereas antibodies against p-eIF2a, eIF2a, and PERK were purchased from Cell Signaling Technology, Inc.

    Techniques: In Vitro, In Vivo, Incubation, Expressing, Western Blot, Mouse Assay, Transfection, Enzyme-linked Immunosorbent Assay

    Expression and activity of Nqo1 in liver cytosolic fractions from wild-type and Nrf2-null mice 1 and 3 days after sham or BDL surgery. (A) Total RNA was isolated from livers of WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously, and analyzed by the bDNA assay for Nqo1 mRNA expression. Data from sham animals at 1 and 3 days were not statistically different and were pooled and denoted as sham. The data are presented as mean relative light units ± standard error of the mean (SEM) ( n = 4–7 animals). (B) Upper panel: Representative Western blot of liver cytosolic fractions from mice 3 days after sham or BDL surgery ( n = 4–7 animals) stained with anti-Nqo1 antibodies. Lower panel: Quantification of Nqo1 protein levels in liver cytosolic fractions from mice 1 and 3 days after sham or BDL surgery. The data are presented as relative protein expression ± SEM ( n = 4–7 animals). (C) Analysis of Nqo1 activity in liver cytosolic fractions from WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously. The data are presented as nanomoles of DCPIP reduced per minute per milligram of protein ± SEM ( n = 4–7 animals). Asterisks (*) represent a statistical difference ( P

    Journal: Cell Stress & Chaperones

    Article Title: Nuclear factor-E2-related factor 2 expression in liver is critical for induction of NAD(P)H:quinone oxidoreductase 1 during cholestasis

    doi: 10.1379/CSC-217.1

    Figure Lengend Snippet: Expression and activity of Nqo1 in liver cytosolic fractions from wild-type and Nrf2-null mice 1 and 3 days after sham or BDL surgery. (A) Total RNA was isolated from livers of WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously, and analyzed by the bDNA assay for Nqo1 mRNA expression. Data from sham animals at 1 and 3 days were not statistically different and were pooled and denoted as sham. The data are presented as mean relative light units ± standard error of the mean (SEM) ( n = 4–7 animals). (B) Upper panel: Representative Western blot of liver cytosolic fractions from mice 3 days after sham or BDL surgery ( n = 4–7 animals) stained with anti-Nqo1 antibodies. Lower panel: Quantification of Nqo1 protein levels in liver cytosolic fractions from mice 1 and 3 days after sham or BDL surgery. The data are presented as relative protein expression ± SEM ( n = 4–7 animals). (C) Analysis of Nqo1 activity in liver cytosolic fractions from WT and Nrf2-null mice that underwent sham or BDL surgery 1 or 3 days previously. The data are presented as nanomoles of DCPIP reduced per minute per milligram of protein ± SEM ( n = 4–7 animals). Asterisks (*) represent a statistical difference ( P

    Article Snippet: Immunochemical detection of Nqo1 protein in cytosolic fractions was performed using anti-Nqo1 (ab2346; Novus Biological, Littleton, CO, USA).

    Techniques: Expressing, Activity Assay, Mouse Assay, Isolation, Western Blot, Staining

    Nqo1 protein expression and activity in liver cytosolic fractions from mice 1, 3, and 7 days after sham or BDL surgery. (A) Representative Western blot of liver cytosolic fractions from mice that underwent sham surgery or BDL surgery (days 1, 3, and 7) stained with anti-Nqo1 antibodies. (B) Quantification of Nqo1 protein levels in liver cytosolic fractions from mice that underwent sham or BDL surgery. The data are presented as relative protein expression ± standard error of the mean (SEM) ( n = 5–13 animals). (C) The data are presented as nanomoles of reduced DCPIP per minute per milligram of protein ± SEM ( n = 5–6 animals). Asterisks (*) represent a statistical difference ( P

    Journal: Cell Stress & Chaperones

    Article Title: Nuclear factor-E2-related factor 2 expression in liver is critical for induction of NAD(P)H:quinone oxidoreductase 1 during cholestasis

    doi: 10.1379/CSC-217.1

    Figure Lengend Snippet: Nqo1 protein expression and activity in liver cytosolic fractions from mice 1, 3, and 7 days after sham or BDL surgery. (A) Representative Western blot of liver cytosolic fractions from mice that underwent sham surgery or BDL surgery (days 1, 3, and 7) stained with anti-Nqo1 antibodies. (B) Quantification of Nqo1 protein levels in liver cytosolic fractions from mice that underwent sham or BDL surgery. The data are presented as relative protein expression ± standard error of the mean (SEM) ( n = 5–13 animals). (C) The data are presented as nanomoles of reduced DCPIP per minute per milligram of protein ± SEM ( n = 5–6 animals). Asterisks (*) represent a statistical difference ( P

    Article Snippet: Immunochemical detection of Nqo1 protein in cytosolic fractions was performed using anti-Nqo1 (ab2346; Novus Biological, Littleton, CO, USA).

    Techniques: Expressing, Activity Assay, Mouse Assay, Western Blot, Staining

    Total hepatic RNA was isolated from mice that underwent sham or BDL surgery at days 1, 3, and 7 and analyzed by the bDNA assay for Nqo1, Ho-1, Gst a1, a4, m1, m2, m3 mRNA expression. The data are presented as mean relative light units ± standard error of the mean ( n = 3–6 animals). Asterisks (*) represent a statistical difference ( P

    Journal: Cell Stress & Chaperones

    Article Title: Nuclear factor-E2-related factor 2 expression in liver is critical for induction of NAD(P)H:quinone oxidoreductase 1 during cholestasis

    doi: 10.1379/CSC-217.1

    Figure Lengend Snippet: Total hepatic RNA was isolated from mice that underwent sham or BDL surgery at days 1, 3, and 7 and analyzed by the bDNA assay for Nqo1, Ho-1, Gst a1, a4, m1, m2, m3 mRNA expression. The data are presented as mean relative light units ± standard error of the mean ( n = 3–6 animals). Asterisks (*) represent a statistical difference ( P

    Article Snippet: Immunochemical detection of Nqo1 protein in cytosolic fractions was performed using anti-Nqo1 (ab2346; Novus Biological, Littleton, CO, USA).

    Techniques: Isolation, Mouse Assay, Expressing

    Salvianolic acid A (SalA) regulates the protein expression of apoptosis-related proteins, Nrf2 signaling, and MAPK signaling. A: Representative protein bands of Bax, Bcl-2, caspase-3 and caspase-9 expression in control, subarachnoid hemorrhage (SAH), and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. B: Representative protein bands of Nrf2, HO-1 and NQO-1 expression in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. C: Representative protein bands of the proteins and the phosphorylation of the proteins in MAPK signaling in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. Values are represented as mean ± standard deviation (n = 5, each group). #P

    Journal: American Journal of Translational Research

    Article Title: Salvianolic acid A attenuates early brain injury after subarachnoid hemorrhage in rats by regulating ERK/P38/Nrf2 signaling

    doi:

    Figure Lengend Snippet: Salvianolic acid A (SalA) regulates the protein expression of apoptosis-related proteins, Nrf2 signaling, and MAPK signaling. A: Representative protein bands of Bax, Bcl-2, caspase-3 and caspase-9 expression in control, subarachnoid hemorrhage (SAH), and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. B: Representative protein bands of Nrf2, HO-1 and NQO-1 expression in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. C: Representative protein bands of the proteins and the phosphorylation of the proteins in MAPK signaling in control, SAH, and SalA-treated (10 and 50 mg/kg) groups were detected by western blot. Values are represented as mean ± standard deviation (n = 5, each group). #P

    Article Snippet: The membranes were respectively incubated at 4°C for 2 h with a mouse monoclonal antibody against Nuclear factor E2 related factor 2 (Nrf2; 1:500, Abcam), HO-1 (1:200, Santa Cruz), NQO-1 (1:200, Santa Cruz), caspase-3 (1:500, Abcam), caspase-9 (1:500, Abcam), Bax (1:500, Abcam), Bcl-2 (1:500, Abcam), extracellular signal regulated kinases (ERK) 1/2 (1:500, Abcam), c-Jun N-terminal kinase (JNK) 1/2 (1:500, Abcam), p38 (1:500, Abcam), or GAPDH (1:500, Abcam).

    Techniques: Expressing, Western Blot, Standard Deviation

    Polydatin protected BMSCs against H 2 O 2 -induced cell death partly through Nrf 2/ARE pathway. BMSCs were pretreated with polydatin for 2 h and further exposed to H 2 O 2 for 12 h. (a) Effects of polydatin on NQO-1 and the phosphorylation of Nrf 2. (b, c) Quantitative analysis of the blots was shown in panel after being normalized by α -tubulin. (d) Cells were treated with different concentration of brusatol for 24 h. Effects of brusatol on phosphorylation of Nrf 2 were detected by Western blot and (g) the bands were normalized by α -tubulin. (e) Cell viability was tested in the presence of different concentration of brusatol. (h) BMSCs were pretreated with brusatol (100 μ M) for 1 h followed by incubating with/without polydatin and H 2 O 2 for 24 h. (f) ROS production was detected by H2DCF-DA staining. (b) Quantitative analysis of DCF fluorescent intensity. One-way ANOVA followed by Tukey's test. ∗ p

    Journal: Stem Cells International

    Article Title: Polydatin Protects Bone Marrow Stem Cells against Oxidative Injury: Involvement of Nrf 2/ARE Pathways

    doi: 10.1155/2016/9394150

    Figure Lengend Snippet: Polydatin protected BMSCs against H 2 O 2 -induced cell death partly through Nrf 2/ARE pathway. BMSCs were pretreated with polydatin for 2 h and further exposed to H 2 O 2 for 12 h. (a) Effects of polydatin on NQO-1 and the phosphorylation of Nrf 2. (b, c) Quantitative analysis of the blots was shown in panel after being normalized by α -tubulin. (d) Cells were treated with different concentration of brusatol for 24 h. Effects of brusatol on phosphorylation of Nrf 2 were detected by Western blot and (g) the bands were normalized by α -tubulin. (e) Cell viability was tested in the presence of different concentration of brusatol. (h) BMSCs were pretreated with brusatol (100 μ M) for 1 h followed by incubating with/without polydatin and H 2 O 2 for 24 h. (f) ROS production was detected by H2DCF-DA staining. (b) Quantitative analysis of DCF fluorescent intensity. One-way ANOVA followed by Tukey's test. ∗ p

    Article Snippet: Further studies showed that polydatin also enhanced phosphorylation of Nrf 2 and upregulation of NQO-1 which was downregulated by H2 O2 , suggesting that polydatin might protect BMSCs against H2 O2 partly via Nrf 2/ARE pathway.

    Techniques: Concentration Assay, Western Blot, Staining

    Antioxidant enzyme synthesis in response to orange oil treatment . Immunoblot analysis demonstrating expression of HO-1, NQO1 and GCLm proteins at 6, 12 and 24 hrs following 15 min treatment of BEAS-2B cells with the oil preparation or time-matched soy oil control. Representative blots from one of three separate experiments are shown above. Densitometric evaluations of each target protein blot normalized to its corresponding GAPDH for all three experiments are provided below. Bars represent mean ± SEM.

    Journal: Respiratory Research

    Article Title: Antioxidant components of naturally-occurring oils exhibit marked anti-inflammatory activity in epithelial cells of the human upper respiratory system

    doi: 10.1186/1465-9921-12-92

    Figure Lengend Snippet: Antioxidant enzyme synthesis in response to orange oil treatment . Immunoblot analysis demonstrating expression of HO-1, NQO1 and GCLm proteins at 6, 12 and 24 hrs following 15 min treatment of BEAS-2B cells with the oil preparation or time-matched soy oil control. Representative blots from one of three separate experiments are shown above. Densitometric evaluations of each target protein blot normalized to its corresponding GAPDH for all three experiments are provided below. Bars represent mean ± SEM.

    Article Snippet: Following antibodies were used for immunoblotting: anti-HO1 (Abcam), anti-NQO1 (Novus Biologicals), anti-GCLm, and anti-GAPDH (Imgenex, Sorrento Valley, CA), anti-Nrf2 and anti-lamin B (Santa Cruze Biotechnology (Santa Cruze, CA).

    Techniques: Expressing

    Kinetics of orange oil-induced antioxidant gene expression . Expression patterns of antioxidant genes HO-1 , NQO1 , GCLm , and GCLc in BEAS-2B cells at designated times following the 15 min treatment period. Data are presented as fold change from time-matched soy oil-treated controls after normalization to the expression of actin. Presented are results from two separate experiments. The dashed line indicates the 2-fold level of increased expression. Results are expressed as mean ± SEM. * indicates significantly different from time-matched soy oil-treated controls (P

    Journal: Respiratory Research

    Article Title: Antioxidant components of naturally-occurring oils exhibit marked anti-inflammatory activity in epithelial cells of the human upper respiratory system

    doi: 10.1186/1465-9921-12-92

    Figure Lengend Snippet: Kinetics of orange oil-induced antioxidant gene expression . Expression patterns of antioxidant genes HO-1 , NQO1 , GCLm , and GCLc in BEAS-2B cells at designated times following the 15 min treatment period. Data are presented as fold change from time-matched soy oil-treated controls after normalization to the expression of actin. Presented are results from two separate experiments. The dashed line indicates the 2-fold level of increased expression. Results are expressed as mean ± SEM. * indicates significantly different from time-matched soy oil-treated controls (P

    Article Snippet: Following antibodies were used for immunoblotting: anti-HO1 (Abcam), anti-NQO1 (Novus Biologicals), anti-GCLm, and anti-GAPDH (Imgenex, Sorrento Valley, CA), anti-Nrf2 and anti-lamin B (Santa Cruze Biotechnology (Santa Cruze, CA).

    Techniques: Expressing

    Treatment of cells with the mixed oil preparation increases expression of oxidant-protective pathways with differing activation kinetics . Time-courses of expression of antioxidant genes HO-1 , NQO1 , GCLm , and GCLc in cells of the BEAS-2B human bronchial epithelial line at designated times following the 15 min treatment period. Data are presented as fold change from time-matched soy oil controls after normalization to the expression of actin. Presented are results from three separate experiments. The dashed line indicates the 2-fold level of increased expression as a reference for potential biological significance. Results are expressed as mean ± SEM. * indicates significantly different from time-matched soy oil controls (P

    Journal: Respiratory Research

    Article Title: Antioxidant components of naturally-occurring oils exhibit marked anti-inflammatory activity in epithelial cells of the human upper respiratory system

    doi: 10.1186/1465-9921-12-92

    Figure Lengend Snippet: Treatment of cells with the mixed oil preparation increases expression of oxidant-protective pathways with differing activation kinetics . Time-courses of expression of antioxidant genes HO-1 , NQO1 , GCLm , and GCLc in cells of the BEAS-2B human bronchial epithelial line at designated times following the 15 min treatment period. Data are presented as fold change from time-matched soy oil controls after normalization to the expression of actin. Presented are results from three separate experiments. The dashed line indicates the 2-fold level of increased expression as a reference for potential biological significance. Results are expressed as mean ± SEM. * indicates significantly different from time-matched soy oil controls (P

    Article Snippet: Following antibodies were used for immunoblotting: anti-HO1 (Abcam), anti-NQO1 (Novus Biologicals), anti-GCLm, and anti-GAPDH (Imgenex, Sorrento Valley, CA), anti-Nrf2 and anti-lamin B (Santa Cruze Biotechnology (Santa Cruze, CA).

    Techniques: Expressing, Activation Assay

    Effect of MitoQ on the Nrf2 signalling in compression‐exposed human NP cells. (A‐J) Human NP cells were treated with or without MitoQ (500 nmol/L) for 12 h. (A‐C) The protein levels of Nrf2 and Keap1 in the human NP cells were measured by Western blotting. (D‐F) The protein expression of nucleus Nrf2 (Nu‐Nrf2) and cytoplasmic Nrf2 (Cyto‐Nrf2) was measured by Western blotting. (G) The nuclear translocation of Nrf2 was examined by immunofluorescence on confocal microscope. Scale bar: 10 μm. (H‐K) The protein levels of SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. (L‐V) Human NP cells were pre‐treated with MitoQ (500 nmol/L) for 2 h and then exposed to compression for 36 h. (L‐Q) The protein levels of Nrf2, Keap1, Nu‐Nrf2 and cyto‐Nrf2 in the human NP cells were measured by Western blotting. (R) The nuclear translocation of Nrf2 was examined by immunofluorescence on confocal microscope. Scale bar: 10 μm. (S‐V) The protein levels of SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. Data are represented as the mean ± SD. *** P

    Journal: Cell Proliferation

    Article Title: The mitochondria‐targeted anti‐oxidant MitoQ protects against intervertebral disc degeneration by ameliorating mitochondrial dysfunction and redox imbalance. The mitochondria‐targeted anti‐oxidant MitoQ protects against intervertebral disc degeneration by ameliorating mitochondrial dysfunction and redox imbalance

    doi: 10.1111/cpr.12779

    Figure Lengend Snippet: Effect of MitoQ on the Nrf2 signalling in compression‐exposed human NP cells. (A‐J) Human NP cells were treated with or without MitoQ (500 nmol/L) for 12 h. (A‐C) The protein levels of Nrf2 and Keap1 in the human NP cells were measured by Western blotting. (D‐F) The protein expression of nucleus Nrf2 (Nu‐Nrf2) and cytoplasmic Nrf2 (Cyto‐Nrf2) was measured by Western blotting. (G) The nuclear translocation of Nrf2 was examined by immunofluorescence on confocal microscope. Scale bar: 10 μm. (H‐K) The protein levels of SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. (L‐V) Human NP cells were pre‐treated with MitoQ (500 nmol/L) for 2 h and then exposed to compression for 36 h. (L‐Q) The protein levels of Nrf2, Keap1, Nu‐Nrf2 and cyto‐Nrf2 in the human NP cells were measured by Western blotting. (R) The nuclear translocation of Nrf2 was examined by immunofluorescence on confocal microscope. Scale bar: 10 μm. (S‐V) The protein levels of SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. Data are represented as the mean ± SD. *** P

    Article Snippet: The membranes were blocked with 7.5% non‐fat milk for 1 hour and then incubated at 4°C overnight with the following primary antibodies: cytochrome c (ab133504, Abcam), cleaved caspase‐3 (#14220, Cell Signaling Technology), Drp1 (ab56788, Abcam), Mff (#84580, Cell Signaling Technology), Fis1 (ab71498, Abcam), Mid49 (bs‐12633R, Bioss), Mid51 (bs‐12634R, Bioss), Mfn1 (#14739, Cell Signaling Technology), Mfn2 (#9482, Cell Signaling Technology), Opa1 (ab42364, Abcam), Parkin (ab77924, Abcam), PINK1 (ab23707, Abcam), p62 (ab56416, Abcam), LC3 (#12741, Cell Signaling Technology), Beclin‐1 (#3495, Cell Signaling Technology), Nrf2 (ab137550, Abcam), Keap1 (10503‐2‐AP, Proteintech), SOD‐2 (24127‐1‐AP, Proteintech), HO‐1 (10701‐1‐AP, Proteintech) and NQO‐1 (11451‐1‐AP, Proteintech).

    Techniques: Western Blot, Expressing, Translocation Assay, Immunofluorescence, Microscopy

    Role of Nrf2 signalling in MitoQ‐mediated anti‐oxidative stress and anti‐mitochondrial disorder in compression‐exposed human NP cells. Human NP cells were transfected with siNrf2 (100 nmol/L) for 48 h followed by the administration of MitoQ and compression. (A‐B) The protein levels of Nrf2, SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. (C‐D) The intracellular ROS levels in the human NP cells were detected using the DCFH‐DA and measured by flow cytometry. (E‐F) The mitochondrial ROS levels were detected using the MitoSOX Red and measured by flow cytometry. (G‐H) Mitochondrial membrane potential was detected by JC‐1 staining and measured by flow cytometry. (I‐J) Annexin V‐APC/7‐AAD staining results showing the rate of apoptosis in human NP cells. Data are represented as the mean ± SD. *** P

    Journal: Cell Proliferation

    Article Title: The mitochondria‐targeted anti‐oxidant MitoQ protects against intervertebral disc degeneration by ameliorating mitochondrial dysfunction and redox imbalance. The mitochondria‐targeted anti‐oxidant MitoQ protects against intervertebral disc degeneration by ameliorating mitochondrial dysfunction and redox imbalance

    doi: 10.1111/cpr.12779

    Figure Lengend Snippet: Role of Nrf2 signalling in MitoQ‐mediated anti‐oxidative stress and anti‐mitochondrial disorder in compression‐exposed human NP cells. Human NP cells were transfected with siNrf2 (100 nmol/L) for 48 h followed by the administration of MitoQ and compression. (A‐B) The protein levels of Nrf2, SOD‐2, HO‐1 and NQO‐1 in the human NP cells were measured by Western blotting. (C‐D) The intracellular ROS levels in the human NP cells were detected using the DCFH‐DA and measured by flow cytometry. (E‐F) The mitochondrial ROS levels were detected using the MitoSOX Red and measured by flow cytometry. (G‐H) Mitochondrial membrane potential was detected by JC‐1 staining and measured by flow cytometry. (I‐J) Annexin V‐APC/7‐AAD staining results showing the rate of apoptosis in human NP cells. Data are represented as the mean ± SD. *** P

    Article Snippet: The membranes were blocked with 7.5% non‐fat milk for 1 hour and then incubated at 4°C overnight with the following primary antibodies: cytochrome c (ab133504, Abcam), cleaved caspase‐3 (#14220, Cell Signaling Technology), Drp1 (ab56788, Abcam), Mff (#84580, Cell Signaling Technology), Fis1 (ab71498, Abcam), Mid49 (bs‐12633R, Bioss), Mid51 (bs‐12634R, Bioss), Mfn1 (#14739, Cell Signaling Technology), Mfn2 (#9482, Cell Signaling Technology), Opa1 (ab42364, Abcam), Parkin (ab77924, Abcam), PINK1 (ab23707, Abcam), p62 (ab56416, Abcam), LC3 (#12741, Cell Signaling Technology), Beclin‐1 (#3495, Cell Signaling Technology), Nrf2 (ab137550, Abcam), Keap1 (10503‐2‐AP, Proteintech), SOD‐2 (24127‐1‐AP, Proteintech), HO‐1 (10701‐1‐AP, Proteintech) and NQO‐1 (11451‐1‐AP, Proteintech).

    Techniques: Transfection, Western Blot, Flow Cytometry, Staining

    Effects of C66 on aortic Nrf2 function. Aortic expression of p-Nrf2 was examined by immunohistochemical staining (A), followed semi-quantitative analysis (B), and Nrf2-downstream genes CAT (C) and NQO-1 (D) expression was examined by real-time PCR at mRNA level. Data were presented as means ± SD ( n = 5); * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Inhibition of JNK by compound C66 prevents pathological changes of the aorta in STZ-induced diabetes

    doi: 10.1111/jcmm.12267

    Figure Lengend Snippet: Effects of C66 on aortic Nrf2 function. Aortic expression of p-Nrf2 was examined by immunohistochemical staining (A), followed semi-quantitative analysis (B), and Nrf2-downstream genes CAT (C) and NQO-1 (D) expression was examined by real-time PCR at mRNA level. Data were presented as means ± SD ( n = 5); * P

    Article Snippet: Primers of JNK, TNF-α, Nrf2, NQO-1 and CAT were purchased from Applied Biosystems (Carlsbad, CA, USA).

    Techniques: Expressing, Immunohistochemistry, Staining, Real-time Polymerase Chain Reaction