Journal: Oxidative Medicine and Cellular Longevity

Article Title: Maggot Extracts Alleviate Inflammation and Oxidative Stress in Acute Experimental Colitis via the Activation of Nrf2

doi: 10.1155/2019/4703253

Figure Lengend Snippet: Sequences of primers designed for RT-qPCR.

Article Snippet: The Nrf2 inhibitor ML385 was purchased from MedChem Express (MCE Co. Ltd., Shanghai, China).

Techniques: Sequencing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Maggot Extracts Alleviate Inflammation and Oxidative Stress in Acute Experimental Colitis via the Activation of Nrf2

doi: 10.1155/2019/4703253

Figure Lengend Snippet: The maggot extracts are capable of suppressing inflammation and oxidative stress in LPS-stimulated RAW 264.7 cells via activation of Nrf2 in a dose-dependent manner. (a) Cells were treated with the maggot extracts for 4 h at various doses as indicated followed by analysis of the induction of Nrf2 and its downstream protein HO-1 with western blotting. GAPDH was used as an internal reference. (b) Cells were treated with a combination of LPS, the maggot extracts (4 μ g/ μ l), or ML385 (3 μ M) as indicated followed by analysis of I κ B, NF κ B p65, IL6, IL-1 β , TNF- α , Nrf2, HO-1, Keap1, p22-phox, and gp91-phox by western blotting. GAPDH was used as an internal reference. Representative data are shown. (c–l) Western blot bands shown in (b) were analyzed by densitometry. The data are shown as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Nrf2 inhibitor ML385 was purchased from MedChem Express (MCE Co. Ltd., Shanghai, China).

Techniques: Activation Assay, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Maggot Extracts Alleviate Inflammation and Oxidative Stress in Acute Experimental Colitis via the Activation of Nrf2

doi: 10.1155/2019/4703253

Figure Lengend Snippet: The maggot extracts prevent DSS-induced experimental colitis in mice. (a) Mice were exposed to drinking water containing 3% DSS for 7 days, followed by normal water for 5 days. Mice were given an intragastric (i.g.) administration of vehicle control, the maggot extracts (1 g/kg), mesalazine (200 mg/kg), or ML385 (30 mg/kg) from day 1 to day 12. Body weight (b), macroscopic appearances (c), colon length (d), and disease activity index (e) were measured and calculated. The data are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Nrf2 inhibitor ML385 was purchased from MedChem Express (MCE Co. Ltd., Shanghai, China).

Techniques: Control, Activity Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Maggot Extracts Alleviate Inflammation and Oxidative Stress in Acute Experimental Colitis via the Activation of Nrf2

doi: 10.1155/2019/4703253

Figure Lengend Snippet: The maggot extracts reduce oxidative stress and inflammation in mice with DSS-induced colitis via activation of the Nrf2 pathway. Mice were treated as explained in . Colonic levels of SOD (a), MPO (b), GSH-Px (c), and MDA (d) were examined by chemical chromatometry. Serum levels of proinflammatory cytokine including TNF- α (e), IL-6 (f), and IL-10 (g) were measured with ELISA. Data are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001; ns = no significance.

Article Snippet: The Nrf2 inhibitor ML385 was purchased from MedChem Express (MCE Co. Ltd., Shanghai, China).

Techniques: Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Maggot Extracts Alleviate Inflammation and Oxidative Stress in Acute Experimental Colitis via the Activation of Nrf2

doi: 10.1155/2019/4703253

Figure Lengend Snippet: The maggot extracts exhibit anti-inflammatory and antioxidant activities in DSS-induced colitis by activating Nrf2. Mice were treated as explained in . (a–j) The expression of p-I κ B, NF κ B p65, IL-6, IL-1 β , TNF- α , p22-phox, gp91-phox, HO-1, Nrf2, and Keap1 in colonic tissues was assessed with western blotting. Representative data are displayed (a) and the relative protein intensity of p-I κ B, NF κ B p65, IL-6, IL-1 β , TNF- α , p22-phox, gp91-phox, HO-1, and Nrf2 was normalized to GAPDH (b–k). (l–o) Sections of colonic tissues were immunostained for revealing various molecules (red) as indicated. The slides were counterstained with DAPI (blue), and the images were captured using an inverted fluorescence microscope. Magnification: ×200. Data are expressed as the mean ± SD (b–k). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The Nrf2 inhibitor ML385 was purchased from MedChem Express (MCE Co. Ltd., Shanghai, China).

Techniques: Expressing, Western Blot, Fluorescence, Microscopy

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Maggot Extracts Alleviate Inflammation and Oxidative Stress in Acute Experimental Colitis via the Activation of Nrf2

doi: 10.1155/2019/4703253

Figure Lengend Snippet: The maggot extracts regulate the proinflammatory cytokines and Nrf2 in the colon of mice with DSS-induced colitis. Mice were treated as explained in . mRNA expression levels of the proinflammatory genes including TNF-α (a), IL-6 (b), and IL-1β (c) as well as oxidative stress-related proteins including Nrf2 (d), p22-phox (e), and Ho-1 (f) in colonic tissues were determined by real-time PCR. The housekeeping gene GAPDH was used as a loading control. Data are expressed as the mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001; ns = no significance.

Article Snippet: The Nrf2 inhibitor ML385 was purchased from MedChem Express (MCE Co. Ltd., Shanghai, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Acta Neuropathologica Communications

Article Title: Identifying new biomarkers of aggressive Group 3 and SHH medulloblastoma using 3D hydrogel models, single cell RNA sequencing and 3D OrbiSIMS imaging

doi: 10.1186/s40478-022-01496-4

Figure Lengend Snippet: A combination of chemotherapy and NRF2 inhibition significantly improves long-term treatment of Group 3 models. a A scheme illustrates the course of the long-term drug treatment assay as previously established . After 3 weeks of growth inside the HA hydrogels the SHH cell lines ONS-76 b , DAOY c and the Group 3 cell lines HD-MB-03 d , D458 e were treated four times with either 10 nM vincristine (green), 5 µM ML385 (NRF2 inhibitor; blue) or a combination (red) or vehicle (black) during one week and cell viability was monitored for the following four weeks in the absence of drug/vehicle present anymore. In the p53 wt SHH cell line ONS-76 only the chemotherapeutic reagent vincristine significantly decreases cell viability, while the NRF2 inhibitor (ML385) alone and in combination with vincristine are also effective in the p53mut SHH cell line DAOY. In contrast, in both Group 3 models the combination of vincristine and NRF2 inhibitor significantly reduced cell viability. (mean ± SEM, n = 3; Two-way ANOVA and Dunnett’s post hoc test, * P < 0.05, ** P < 0.01 and *** P < 0.001 all relative to DMSO according to colour code). Analysis of the biggest publicly available MB data base shows that NRF2 (gene name: NFE2L2 ) gene expression does not predict survival of SHH ( f ; logrank test, p = 0.924) patients, but is associated with worse survival in Group 3 patients ( g ; logrank test, p = 0.078). Note the exclusive effect of NRF2 expression in Group 3 patients

Article Snippet: Three-week old gels were treated with either 10 nM vincristine (VCR) (S1241; Selleckchem), 5 µM NRF2 inhibitor ML385 (6243, Bio-Techne) or both for one week.

Techniques: Inhibition, Expressing

Journal: Acta Neuropathologica Communications

Article Title: Identifying new biomarkers of aggressive Group 3 and SHH medulloblastoma using 3D hydrogel models, single cell RNA sequencing and 3D OrbiSIMS imaging

doi: 10.1186/s40478-022-01496-4

Figure Lengend Snippet: HR-MAS NMR spectroscopy detected fumarate levels group 3 and high LUM gene expression levels in SHH MB patients predict better overall survival. HR-MAS NMR spectroscopy analysis of fumarate in SHH ( a , n = 13) and Group 3 ( b , n = 16) patients indicates no survival difference for SHH patients (no fumarate detected n = 8; fumarate detected n = 5) but a strong trend for worse overall survival in Group 3 patients (no fumarate detected n = 12; fumarate detected n = 4) with detectable fumarate levels. c Genomic analysis of LUM expression in SHH ( c ; logrank test, p < 0.001) and Group 3 ( d ; logrank test, p = 0.056) patients identified that high LUM expression correlates with better overall survival than low LUM expression, whereas the opposite trend is true in Group 3 patients. e A graphical abstract of this study illustrating that by using by using a combination of state-of-the-art techniques (scRNAseq and 3D OrbiSIMS), in a realistic 3D model, we have identified sub-group-specific tumour phenotypes in SHH and Group 3 medulloblastoma. In the SHH sub-group we have demonstrated the formation of an ECM shell-like structure composed of laminin, type I- and VI-collagens and lumican that can be used to identify low-risk SHH tumours. This could facilitate more accurate risk stratification of SHH medulloblastoma. In the Group 3 sub-group we have shown that fumarate accumulation identifies very-high risk patients that could benefit from a chemotherapy combination with NRF2 targeted therapy approaches

Article Snippet: Three-week old gels were treated with either 10 nM vincristine (VCR) (S1241; Selleckchem), 5 µM NRF2 inhibitor ML385 (6243, Bio-Techne) or both for one week.

Techniques: Spectroscopy, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of LPS on NRF-2 activation in NRK-52e cells. A) Cell viability after addition of 6–50 μg/mL LPS for 24 h (n=5). B) Cell viability after addition of 50 μg/mL LPS for 6–24 h (n=5). C) Western blots showing NRF2 expression in cells at 6, 12, and 24 h after addition of LPS (50 μg/mL). D) Quantitative analysis of the western blot data, with expression relative to β-actin (n=3). E) Immunofluorescence detection of the expression and distribution of NRF2 following LPS treatment. Data are presented as means ± SD; *p<0.05 vs control.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Activation Assay, Western Blot, Expressing, Immunofluorescence

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of NRF2 inhibition on viability and apoptosis in LPS-induced NRK-52e cells. A) Representative images of apoptotic cells measured by the TUNEL assay. B) Cell viability, determined by the MTT assay (n=5). (C) Western blotting of Bcl-2, Bax, cytochrome c, and NRF2. D-I) Quantitative analysis of the Western blotting data, with expression relative to β-actin, Lamin BI, or COXIV (n=3). Data are presented as the means ± SD; *p<0.05 vs control, # p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Inhibition, TUNEL Assay, MTT Assay, Western Blot, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of NRF2 on the inflammatory response and oxidative stress in LPS-induced injury of NRK-52e cells. A) Immunoblotting of NRF2 following transfection with LV-NRF2. B) Western blotting of NRF2, Nucleus NRF2, p-p65, IKB-α, p-p38 and erk. C) Quantitative analysis of the Western blotting data, with expression relative to β-actin, p65, p38 or Lamin B1 (n=3). D,E) ELISA of TNF-α and IL-6 in cell culture supernatants (n=5). (F) SOD activity (n=5). G) MDA level (n=5). H) Representative fluorescence microscopy images of ROS production (DCFH-DA probe). I) Quantitative analysis of the ROS data (n=5). Data are presented as means ± SD; *p<0.05 vs control, # p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Western Blot, Transfection, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Activity Assay, Fluorescence, Microscopy

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of NRF2 inhibition on viability and apoptosis in LPS-induced NRK-52e cells. A) Representative images of apoptotic cells measured by the TUNEL assay. B) Cell viability, determined by the MTT assay (n=5). (C) Western blotting of Bcl-2, Bax, cytochrome c, and NRF2. D-I) Quantitative analysis of the Western blotting data, with expression relative to β-actin, Lamin BI, or COXIV (n=3). Data are presented as the means ± SD; *p<0.05 vs control, # p<0.05 vs LPS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Inhibition, TUNEL Assay, MTT Assay, Western Blot, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of NRF2 on mitochondrial homeostasis of LPS-induced NRK-52e cells. A) Western blotting of proteins related to mitophagy and mitochondrial-biogenesis. B-F) Quantitative analysis of the Western blotting data, with expression relative to β-actin (n=3). G) Representative fluorescence microscopy images showing co-localization of anti-LC3 II (green)/anti-COXIV (red) antibodies. Data are presented as means ± SD; *p<0.05 vs control, # p<0.05 vs L PS.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Western Blot, Expressing, Fluorescence, Microscopy

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of CLP of rats on renal expression of NRF2. A) Western blotting of NRF2 in kidney tissues. B) Quantitative analysis of the Western blotting data, with expression relative to β-actin (n=5). C) Representative immunohistochemical images showing NRF2 expression. Data are presented as means ±SD; *p<0.05 vs control.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Expressing, Western Blot, Immunohistochemical staining

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of NRF2 activation on oxidative stress, inflammatory responses, and renal function in rats that received CLP. A,B) Western blotting of NRF2 in kidney tissues. C-E) Quantitative analysis Western blotting data, with expression relative to β-actin or Lamin B1 (n=5). F) MDA levels in kidney tissues (n=8). G) SOD activity in kidney tissues (n=8). H,I) ELISA of serum levels TNF-α and IL-6 (n=8). J,K) Serum creatinine and BUN (n=8). L) Representative hematoxylin and eosin staining images of renal tissues. Data are presented as means ± SDs; asterisks, edema; triangle, vacuolization; arrow, loss of brush border; *p<0.05 vs sham; #p<0.05 vs CLP.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Activation Assay, Western Blot, Expressing, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: European Journal of Histochemistry : EJH

Article Title: Transcription factor nuclear factor erythroid 2 p45-related factor 2 (NRF2) ameliorates sepsis-associated acute kidney injury by maintaining mitochondrial homeostasis and improving the mitochondrial function

doi: 10.4081/ejh.2022.3412

Figure Lengend Snippet: Effect of NRF2 on mitochondrial damage and mitochondrial homeostasis in rats that received CLP. A) Western blotting of kidney proteins related to mitophagy and mitochondrial biogenesis. B-F) Quantitative analysis of the western blotting data with expression relative to β-actin (n=5; data are presented as means ± SD; *p<0.05 vs sham, # p<0.05 v s CLP). G) ATP level in kidney tissues (n=8). H) Representative transmission electron micrographs of renal mitochondria. M, mitochondrion; N, nucleus; CLP, the arrow shows a mitochondrion with swelling and cristae fracture; CLP+ML385, the arrow shows a mitochondrion with swelling, cristae fracture, and rupture of membranes; CLP+TBHQ, the arrow shows a mitochondrion with mild swelling and cristae fracture.

Article Snippet: The dose of NRF2 inhibitor and NRF2 agonist were selected based on previous reports., The NRF2 inhibitor ML385 (T4360, TargetMol Chemicals Inc., Boston, MA, USA) was administered by intraperitoneal injection (30 mg/kg) at 4 h before the CLP procedure in the CLP 24 h + ML385 group; the NRF2 agonist tert-butylhydroquinone (TBHQ) (HY-100489, MedChemExpress LLC, Princeton, NJ, USA) was administered by intraperitoneal injection (50 mg/kg) at 4 h before CLP in the CLP 24 h + TBHQ group.

Techniques: Western Blot, Expressing, Transmission Assay

Journal: Marine Drugs

Article Title: Fucoxanthin Pretreatment Ameliorates Visible Light-Induced Phagocytosis Disruption of RPE Cells under a Lipid-Rich Environment via the Nrf2 Pathway

doi: 10.3390/md20010015

Figure Lengend Snippet: The effect of pretreatment time on Nrf2 signaling pathway activated by fucoxanthin in differentiated RPE cells. ( a ) Nucl-Nrf2 activity; ( b ) GCLC expression level; ( c ) GPx expression level; ( d ) TrxR expression level; ( e ) HO-1 expression level; ( f ) NQO1 expression level. (** p < 0.01 vs. control; ▲▲ p < 0.01 vs. lutein).

Article Snippet: The Nrf2 inhibitor ML385 was provided by Sigma-Aldrich (MO).

Techniques: Activity Assay, Expressing

Journal: Marine Drugs

Article Title: Fucoxanthin Pretreatment Ameliorates Visible Light-Induced Phagocytosis Disruption of RPE Cells under a Lipid-Rich Environment via the Nrf2 Pathway

doi: 10.3390/md20010015

Figure Lengend Snippet: Ameliorative effects of fucoxanthin on phagocytosis disorder in RPE cells via the Nrf2 signal pathway: ( a ) the expressions of Nucl-Nrf2, NQO1, and HO-1 when RPE cells were treated with fucoxanthin or fucoxanthin +ML385; ( b ) ROS production when RPE cells were treated with fucoxanthin or fucoxanthin + ML385; ( c ) TNF-α levels when RPE cells were treated with fucoxanthin or fucoxanthin +ML385; ( d ) phagocytic indexes when RPE cells were treated with fucoxanthin or fucoxanthin +ML385. (* p < 0.05 and ** p < 0.01 vs. control; ## p < 0.01 vs. light exposure).

Article Snippet: The Nrf2 inhibitor ML385 was provided by Sigma-Aldrich (MO).

Techniques:

Journal: Cells

Article Title: Activation of Nrf2 in Astrocytes Suppressed PD-Like Phenotypes via Antioxidant and Autophagy Pathways in Rat and Drosophila Models.

doi: 10.3390/cells10081850

Figure Lengend Snippet: Figure 2. CDDO-Me activated Nrf2/ARE pathway predominantly in astrocytes in 6-OHDA rat brains. (A) Nrf2 immunos- taining. Representative pictures show Nrf2 immunofluorescence in SNpc at week 2 after 6-OHDA injection, and nuclei were re-stained with Hoechst (200× magnification). The bar graph on the right side of the pictures is the quantitative analysis of optical density for fluorescence. The Nrf2-positive cells were few in the control group and mainly distributed in the cytoplasm. The Nrf2 signal increased in the 6-OHDA group compared with the control group (* p < 0.05, n = 5) and was mainly distributed in the nucleus. CDDO-Me increased Nrf2-positive cells in the 6-OHDA + CDDO group compared with the 6-OHDA group and control group (* p < 0.05,** p < 0.01, n = 5), and the Nrf2 signal was also mainly distributed in the nucleus in the 6-OHDA + CDDO group. (B) Co-immunostaining of Nrf2 with GFAP. Representative pictures show Nrf2 fluorescence was mainly distributed in neurons but not in GFAP-positive astrocytes in control group. Increased Nrf2 signal was colocalized with GFAP staining in the 6-OHDA group and 6-OHDA + CDDO group. The bar graph on the right side of the pictures is the quantitative analysis of optical density for Nrf2 fluorescence colocalization with GFAP-positive astrocytes. The results show more Nrf2 signal colocalized with GFAP fluorescence in the 6-OHDA group compared with the control group (* p < 0.05, n = 5). CDDO-Me further increased the Nrf2-positive signal in GFAP-positive astrocytes in the 6-OHDA + CDDO group compared with the 6-OHDA group (* p < 0.05, ** p < 0.01, n = 5). (C) Representative Western blots of Nrf2 and GCLC proteins in the substantia nigra and striatum homogenate. The bar graph below is the quantitative analysis of Western blot bands. Nrf2 did not significantly change in the substantia nigra (NS p > 0.05) and increased in the striatum in the 6-OHDA group compared with the control group (* p < 0.05). CDDO-Me increased Nrf2 in both substantia nigra and striatum in the 6-OHDA + CDDO group compared with in the 6-OHDA group (* p < 0.05). The Nrf2 downstream antioxidant GCLC also significantly increased in the substantia nigra and striatum in the 6-OHDA group compared with the control group (* p < 0.05). CDDO-Me further increased the GCLC level in the 6-OHDA + CDDO group compared with in the 6-OHDA group (* p < 0.05). (D) Representative Western blots of LC3 and quantitative analysis. The expression of the autophagy-related protein LC3-II did not significantly change in the substantia nigra and striatum in the 6-OHDA group compared with the control group (NS p > 0.05). CDDO-Me effectively increased LC3-II expression in the 6-OHDA + CDDO group compared with the 6-OHDA group (* p < 0.05).

Article Snippet: The Nrf2 inhibitor ML385 (S8790, Selleck, Houston, TX, USA), Nrf2 inducer CDDOMe (S8078, Selleck), autophagy inhibitor 3-MA (S2767, Selleck), and autophagy inducer rapamycin (S1039, Selleck) were added to the Drosophila food at a final concentration of 10 μM.

Techniques: Injection, Staining, Control, Immunostaining, Western Blot, Expressing

Journal: Cells

Article Title: Activation of Nrf2 in Astrocytes Suppressed PD-Like Phenotypes via Antioxidant and Autophagy Pathways in Rat and Drosophila Models.

doi: 10.3390/cells10081850

Figure Lengend Snippet: Figure 4. Overexpression of Nrf2 in glial cells suppressed rotenone-induced PD-like phenotypes in flies. (A) survival curve and statistical analysis by Log-rank test and half survival time. Nrf2 overexpression in DMSO control drosophila did not significantly improve survival time in the DMSO/Elav;cncc or DMSO/Repo;cncc group compared with the DMSO/WT control group (NS p > 0.05). Rotenone treatment significantly shortened survival time compared with the corresponding DMSO control group (## p < 0.01). Nrf2 overexpression in neurons of PD model Drosophila significantly increased survival time in the rotenone/Elav;cncc group compared with the rotenone/WT group (* p < 0.05). Nrf2 overexpression in glial cells more significantly increased survival time in the rotenone/Repo;cncc group compared with the rotenone/Elav;cncc group and the rotenone/WT group (* p < 0.05 and ** p < 0.01). (B) The left graph shows climbing ability throughout lifetime. The right bar graph shows the climbing ability at ages of 3–5 weeks. Rotenone treatment significantly reduced climbing ability in the rotenone/WT group compared with the DMSO group (## p < 0.01). Nrf2 overexpression in glial cells in the rotenone/Repo;cncc group more significantly improved the motor ability than overexpression in neurons in the rotenone/Elav;cncc group (* p < 0.05 and ** p < 0.01). (C) Western blot analysis of Nrf2, GCLC, HO-1, and Ref(2)P proteins in Drosophila brain homogenates. The bar graph below is the quantitative analysis of Western blot bands. Nrf2 overexpression in neurons and glial cells increased GCLC and HO-1 in the Elav;cncc and Repo;cncc group compared with the WT control group (* p < 0.05 and ** p < 0.01). Nrf2 overexpression in glial cells further increased Nrf2, GCLC and HO-1 in the Repo;cncc group compared to overexpression in neurons in the Elav;cncc group (* p < 0.05). The Nrf2 overexpression significantly increased Ref(2)P in the rotenone/Elav;cncc and rotenone/Repo;cncc groups compared with the rotenone/WT

Article Snippet: The Nrf2 inhibitor ML385 (S8790, Selleck, Houston, TX, USA), Nrf2 inducer CDDOMe (S8078, Selleck), autophagy inhibitor 3-MA (S2767, Selleck), and autophagy inducer rapamycin (S1039, Selleck) were added to the Drosophila food at a final concentration of 10 μM.

Techniques: Over Expression, Control, Western Blot

Journal: Cells

Article Title: Activation of Nrf2 in Astrocytes Suppressed PD-Like Phenotypes via Antioxidant and Autophagy Pathways in Rat and Drosophila Models.

doi: 10.3390/cells10081850

Figure Lengend Snippet: Figure 5. Activation of the Nrf2 signaling pathway enhanced autophagy in flies. (A) The graph shows the lifetime survival curve, the Log-rank test, and half survival time used to analyze the difference between groups (#, compared with the rotenone/WT group). Nrf2 overexpression in glial cells effectively prolonged survival time in the Rot/Repo;cncc + DMSO group compared with the rotenone/WT group (## p < 0.01). The Nrf2 inhibitor ML385 significantly reduced the prolonged survival time in the Rot/Repo;cncc + ML385 group compared with the Rot/Repo;cncc + DMSO group (** p < 0.01). The autophagy inhibitor 3-MA partly reduced the prolonged survival time of the Rot/Repo;cncc + 3-MA group (* p < 0.05). The Nrf2 inducer CDDO-Me and autophagy inducer rapamycin did not significantly affect the survival time of the Rot/Repo;cncc + DMSO group. (B) Western blot analysis of antioxidant and autophagy proteins in fly brain. Nrf2 overexpression in glial cells significantly increased HO-1 and Ref(2)P proteins in the rotenone/Repo;cncc group compared with the rotenone/WT group (## p < 0.01). The Nrf2 inhibitor ML385 effectively decreased HO-1 and Ref(2)P levels in the rotenone/Repo;cncc + ML385 group compared with the rotenone/Repo;cncc + DMSO group (* p < 0.05). The autophagy inhibitor 3-MA effectively decreased Ref(2)P protein but not HO-1 protein, while the Nrf2 inducer CDDO-Me and autophagy inducer rapamycin did not significantly affect the expression of HO-1 and Ref(2)P in the rotenone/Repo;cncc + DMSO group.

Article Snippet: The Nrf2 inhibitor ML385 (S8790, Selleck, Houston, TX, USA), Nrf2 inducer CDDOMe (S8078, Selleck), autophagy inhibitor 3-MA (S2767, Selleck), and autophagy inducer rapamycin (S1039, Selleck) were added to the Drosophila food at a final concentration of 10 μM.

Techniques: Activation Assay, Over Expression, Western Blot, Expressing

Journal: Cells

Article Title: Activation of Nrf2 in Astrocytes Suppressed PD-Like Phenotypes via Antioxidant and Autophagy Pathways in Rat and Drosophila Models.

doi: 10.3390/cells10081850

Figure Lengend Snippet: Figure 6. Knockdown of Nrf2 in fly glial cells worsens rotenone-induced PD-like phenotypes in flies. (A) The survival curve in Nrf2-knockdown Drosophila. The graph shows the survival curve, the Log-rank test, and half survival time used to analyze the difference between groups (#, compared with the rotenone/WT group). Nrf2 knockdown in glial cells reduced survival time in the Rot/Repo;cnccRNAi + DMSO group compared with the rotenone/WT group (# p < 0.05). CDDO-Me significantly extended survival time in the Rot/Repo;cnccRNAi + CDDO-Me group compared with the Rot/Repo;cnccRNAi + DMSO group (** p < 0.01). Rapamycin partially restored the shortened survival time of the Rot/Repo;cnccRNAi + DMSO group (* p < 0.05). (B) qRT-PCR result for drosophila Nrf2, the cncc level in Repo;cnccRNAi flies was about 40% compared with WT Control (## p < 0.01). (C) Western blot analysis of antioxidant and autophagy proteins in fly brain. Nrf2 knockdown in glial cells by RNAi significantly decreased protein expression of HO-1 and Ref(2)P in the rotenone/Repo;cnccRNAi group compared with the rotenone/WT group (# p < 0.05 and ## p < 0.01). CDDO-Me partially restored this decrease in HO-1 and Ref(2)P expression in the rotenone/Repo;cnccRNAi + CDDO-Me group compared with the rotenone/Repo;cnccRNAi + DMSO group (* p < 0.05). Rapamycin partially restored the expression of the Ref(2)P autophagy protein, but did not alter HO-1 levels in the rotenone/Repo;cnccRNAi + Rapa group compared with the rotenone/Repo;cnccRNAi + DMSO group (* p < 0.05).

Article Snippet: The Nrf2 inhibitor ML385 (S8790, Selleck, Houston, TX, USA), Nrf2 inducer CDDOMe (S8078, Selleck), autophagy inhibitor 3-MA (S2767, Selleck), and autophagy inducer rapamycin (S1039, Selleck) were added to the Drosophila food at a final concentration of 10 μM.

Techniques: Knockdown, Quantitative RT-PCR, Control, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: PMN-MDSCs Enhance CTC Metastatic Properties through Reciprocal Interactions via ROS/Notch/Nodal Signaling

doi: 10.3390/ijms20081916

Figure Lengend Snippet: ( A ) Dose-dependent increase of NRF2 target genes in melanoma CTCs following H 2 O 2 treatment. Data from a representative experiment ( n = 4) are shown. ( B ) Augmented gene expression of Notch1 ligands Jagged1 and DLL4 in PMN-MDSCs isolated from patients with/without brain metastasis (BM). HD = corresponding analyses of PMN-MDSCs isolated from blood of healthy donors. ( C ) CTC proliferation is up-regulated by the combinatorial treatment of oxidative stress and MDSC Jagged1 measured by real-time IncuCyte ® Live-Cell Analysis System in 3D cell conditions. No treatment (negative control; black), H 2 O 2 (25 μM; red), recombinant human Jagged1 (100 μM; yellow), and combinatorial (orange) are shown. Data are representative of six independent experiments ( n = 6).

Article Snippet: NRF2 inhibitor ML385 (#2114) was obtained from Cayman, Inc. while Nodal inhibitor Lefty (746-LF/CF) was purchased from R&D Systems, Inc. (Waltham, MA, USA).

Techniques: Gene Expression, Isolation, Cell Analysis, Negative Control, Recombinant

Journal: International Journal of Molecular Sciences

Article Title: PMN-MDSCs Enhance CTC Metastatic Properties through Reciprocal Interactions via ROS/Notch/Nodal Signaling

doi: 10.3390/ijms20081916

Figure Lengend Snippet: Model outlining biomarkers and pathways between CTCs and PMN-MDSCs interactions (CTC/PMN-MDSC cluster) isolated from blood of metastatic cancer patients. Large red arrow shows PMN-MDSC-secreted ROS affecting CTC-associated Nrf2/Notch1/Nodal pathway. Small black arrows refer to induction of CTC Nrf2-ARE, Notch1 and Nodal gene expression, respectively.

Article Snippet: NRF2 inhibitor ML385 (#2114) was obtained from Cayman, Inc. while Nodal inhibitor Lefty (746-LF/CF) was purchased from R&D Systems, Inc. (Waltham, MA, USA).

Techniques: Isolation, Gene Expression

Journal: Cancers

Article Title: Methods for Overcoming Chemoresistance in Head and Neck Squamous Cell Carcinoma: Keeping the Focus on Cancer Stem Cells, a Systematic Review

doi: 10.3390/cancers16173004

Figure Lengend Snippet: Molecules adjuvant to chemotherapy and their effects on CSCs.

Article Snippet: In a study conducted by Praharaj et al. [ ], the efficacy of ML-385 (a pharmacological inhibitor of NRF2) and chloroquine (CQ) on FaDu and tongue carcinoma Cal-33 cells (DSMZ, ACC447) was evaluated.

Techniques: Adjuvant, Expressing, In Vitro, In Vivo