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: Artificial cells, nanomedicine, and biotechnology

Article Title: Protective effect of ginsenoside Rg1 on attenuating anti-GBM glomerular nephritis by activating NRF2 signalling.

doi: 10.1080/21691401.2019.1640712

Figure Lengend Snippet: Figure 2. Ginsenoside Rg1 attenuated IL1b-induced apoptosis and activated NRF2 signalling. (A) Western blot analysis indicated Ginsenoside Rg1 protected podo- cytes from IL1b-induced apoptosis, (n ¼ 3). (B–D) Ginsenoside Rg1 activated NRF2 signalling and inhibited IL1b-stimulated ERK and P38 pathway (n ¼ 3). p < .05; p < .01; versus respective control.

Article Snippet: Ginsenoside Rg1 (80 ng/mL) was exposed to podocytes co-cultured with recombinant human IL-1b protein (10 ng/mL) (R&D, USA) in presence or absence of NRF2 inhibitor ML385 (5lM,) (Selleck, USA) for 48 h.

Techniques: Western Blot, Control

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Protective effect of ginsenoside Rg1 on attenuating anti-GBM glomerular nephritis by activating NRF2 signalling.

doi: 10.1080/21691401.2019.1640712

Figure Lengend Snippet: Figure 3. NRF2 inhibitor ML385 inhibited NRF2 signalling and promoted IL-1b-induced apoptosis in podocytes. (A–C) ML385 inhibited NRF2 signalling and re-acti- vated ERK and P38 signalling pathway (n ¼ 3). (D) ML385 diminished the inhibitory effect of Ginsenoside Rg1 on attenuating apoptosis in podocytes (n ¼ 3). p < .05; p<.01; versus respective control.

Article Snippet: Ginsenoside Rg1 (80 ng/mL) was exposed to podocytes co-cultured with recombinant human IL-1b protein (10 ng/mL) (R&D, USA) in presence or absence of NRF2 inhibitor ML385 (5lM,) (Selleck, USA) for 48 h.

Techniques: Control

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Protective effect of ginsenoside Rg1 on attenuating anti-GBM glomerular nephritis by activating NRF2 signalling.

doi: 10.1080/21691401.2019.1640712

Figure Lengend Snippet: Figure 4. NRF2 inhibitor ML385 abrogated anti-inflammatory effect of Ginsenoside Rg1 on podocytes. (A–D) ML385 eliminated inhibitory effect of Ginsenoside Rg1 on attenuating inflammation in podocytes (n ¼ 6). p < .05; p < .01; p < .001; versus respective control.

Article Snippet: Ginsenoside Rg1 (80 ng/mL) was exposed to podocytes co-cultured with recombinant human IL-1b protein (10 ng/mL) (R&D, USA) in presence or absence of NRF2 inhibitor ML385 (5lM,) (Selleck, USA) for 48 h.

Techniques: Control

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Protective effect of ginsenoside Rg1 on attenuating anti-GBM glomerular nephritis by activating NRF2 signalling.

doi: 10.1080/21691401.2019.1640712

Figure Lengend Snippet: Figure 6. Ginsenoside Rg1 showed beneficial effect on inhibiting apoptosis and activated NRF2 signalling in mice. (A) Ginsenoside Rg1 protected renal function from apoptosis (n ¼ 3). (B–D) Ginsenoside Rg1 activated NRF2 signalling and inhibited ERK and P38 pathway in vivo (n ¼ 3). p < .05; p < .01; versus respect- ive control.

Article Snippet: Ginsenoside Rg1 (80 ng/mL) was exposed to podocytes co-cultured with recombinant human IL-1b protein (10 ng/mL) (R&D, USA) in presence or absence of NRF2 inhibitor ML385 (5lM,) (Selleck, USA) for 48 h.

Techniques: In Vivo, Control

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates ferroptosis after subarachnoid hemorrhage via Nrf2/HO-1 signaling pathway

doi: 10.3389/fphar.2022.924826

Figure Lengend Snippet: Animal usage and mortality.

Article Snippet: ML385 (50 pmol/5 μl; AOBIOUS, Gloucester, MA, United States), a specific Nrf2 inhibitor, was diluted in DMSO, and was used for intracerebroventricular (i.c.v.) injection at 24 h before SAH.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Astragaloside IV attenuates ferroptosis after subarachnoid hemorrhage via Nrf2/HO-1 signaling pathway

doi: 10.3389/fphar.2022.924826

Figure Lengend Snippet: ML385 inhibits the neuroprotective effect of AS-IV at 24 h post-SAH. (A–C) Modified Garcia scores, beam balance scores, and brain water content 24 h after SAH in rats ( n = 8). (D,E) Representative images and quantitative analysis of FJB staining in right temporal cerebral cortex of rats (n = 6, scale bar = 50 μm). (F) Quantification of iron concentrations in rats after SAH ( n = 8). (G–I) Quantitative analysis of lipid ROS, MDA, and GSH levels ( n = 8). (J) Representative Western blot images of SLC7A11 and GPX4. (K) Quantitative analysis of SLC7A11 and GPX4 ( n = 6). Data are represented as mean ± SD. * p < 0.05, NS: not statistically significant; DMSO: dimethyl sulfoxide; FJB, Fluoro-Jade B .

Article Snippet: ML385 (50 pmol/5 μl; AOBIOUS, Gloucester, MA, United States), a specific Nrf2 inhibitor, was diluted in DMSO, and was used for intracerebroventricular (i.c.v.) injection at 24 h before SAH.

Techniques: Modification, Staining, Western Blot