Journal: Pharmaceutics

Article Title: Myrrh Oil-Based Nanoemulsion Loaded with Curcumin and Insulin: Development, Characterization, and Evaluation of Enhanced Antibacterial and Diabetic Wound-Healing Activity

doi: 10.3390/pharmaceutics18030369

Figure Lengend Snippet: Effect of topical diabetic wound treatments with various preparations on biomarkers of oxidative stress. ( A ): Nuclear factor erythroid 2-related factor-2 (Nrf-2); ( B ): malondialdehyde (MDA); ( C ): reduced glutathione (GSH). ###: Significantly different compared with the normal control group at p < 0.001, #### at p < 0.0001. *: Significantly different compared with DM + B-NEG group at p < 0.05, ** at p < 0.01, *** at p < 0.001, **** at p < 0.0001. $$$$: Significantly different compared with the CUR-G group at p < 0.0001. @: Significantly different compared with INS-G group at p < 0.05, @@ at p < 0.001.

Article Snippet: An enzyme-linked immunosorbent assay (ELISA) kit was used to measure the concentration of nuclear factor erythroid 2-related factor 2 (Nrf-2, BT LAB, Shanghai, China) in skin tissue homogenate following the manufacturer’s procedures.

Techniques: Control

Journal: The Journal of nutrition

Article Title: Ajoene, a stable garlic by-product, has an antioxidant effect through Nrf2-mediated glutamate-cysteine ligase induction in HepG2 cells and primary hepatocytes.

doi: 10.3945/jn.110.121277

Figure Lengend Snippet: FIGURE 1 Activation of Nrf2 by ajoene in HepG2 cells treated with 3–30 mmol/L ajoene or 30 mmol/L t-BHQ for 6 h (A) or 30 mmol/L ajoene for 3–12 h (B) and ARE reporter gene activity in HepG2 cells treated with 3–30 mmol/L ajoene or 30 mmol/L t-BHQ for 24 h after transfection of the NQO1-ARE luciferase construct. Values are means 6 SD, n = 4. Means without a common letter differ, P , 0.05.

Article Snippet: Antibodies directed against phospho-PKCd, phospho-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), phospho-c-Jun NH2-terminal kinase (JNK) (Thr183/ Tyr185), phospho-Akt, and lamin A/C were obtained from Cell Signaling Technology, whereas that of phospho-Nrf2 (Ser40) was supplied from Novus Biologicals.

Techniques: Activation Assay, Activity Assay, Transfection, Luciferase, Construct

Journal: The Journal of nutrition

Article Title: Ajoene, a stable garlic by-product, has an antioxidant effect through Nrf2-mediated glutamate-cysteine ligase induction in HepG2 cells and primary hepatocytes.

doi: 10.3945/jn.110.121277

Figure Lengend Snippet: FIGURE 3 Inhibition of Nrf2 ubiquitination by ajoene in HepG2 cells. (A) The interaction between Keap1 and Nrf2. Immunoprecipitated Nrf2 was subjected to immunoblotting for Keap1. Results were confirmed in 3 replicate experiments. (B) Nrf2 ubiquitination. Cells were transfected with the plasmid encoding His-tagged ubiquitin (His-Ubi), treated with 10 mmol/L MG132 (a proteasomal inhibitor) for 1 h, and continuously incubated with 30 mmol/L ajoene or 30 mmol/L t-BHQ for 6 h. Immunoprecipitated His-Ubi was subjected to immunoblottings for Nrf2. Mock transfection of pcDNA3 (an empty vector) was used as control. Values are means 6 SD, n = 4. Means without a common letter differ, P , 0.05. IP, Immunoprecipitation; IB, immunoblotting.

Article Snippet: Antibodies directed against phospho-PKCd, phospho-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), phospho-c-Jun NH2-terminal kinase (JNK) (Thr183/ Tyr185), phospho-Akt, and lamin A/C were obtained from Cell Signaling Technology, whereas that of phospho-Nrf2 (Ser40) was supplied from Novus Biologicals.

Techniques: Inhibition, Ubiquitin Proteomics, Immunoprecipitation, Western Blot, Transfection, Plasmid Preparation, Incubation, Control

Journal: The Journal of nutrition

Article Title: Ajoene, a stable garlic by-product, has an antioxidant effect through Nrf2-mediated glutamate-cysteine ligase induction in HepG2 cells and primary hepatocytes.

doi: 10.3945/jn.110.121277

Figure Lengend Snippet: FIGURE 4 PKCd-dependent increases in GCLC and GCLM mRNA levels in HepG2 cells. (A) The effects of ajoene on the phosphorylation of upstream kinases. Immunoblot analyses were performed on cell lysates that had been incubated with 30 mmol/L ajoene for 10 min to 6 h. (B) Inhibition of Nrf2 phosphorylation and nuclear accumulation by rottlerin. Cells were treated with 30 mmol/L ajoene alone or in combination with 2 mmol/L rottlerin or 5 mmol/L U73122 for 6 h. Results were confirmed in 3 replicate experiments. GCLC (C) and GCLM (D) mRNA levels measured using real-time PCR assays in cells that had been treated as described in B. Values are means 6 SD, n = 4. Means without a common letter differ, P , 0.05.

Article Snippet: Antibodies directed against phospho-PKCd, phospho-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), phospho-c-Jun NH2-terminal kinase (JNK) (Thr183/ Tyr185), phospho-Akt, and lamin A/C were obtained from Cell Signaling Technology, whereas that of phospho-Nrf2 (Ser40) was supplied from Novus Biologicals.

Techniques: Phospho-proteomics, Western Blot, Incubation, Inhibition, Real-time Polymerase Chain Reaction

Journal: The Journal of nutrition

Article Title: Ajoene, a stable garlic by-product, has an antioxidant effect through Nrf2-mediated glutamate-cysteine ligase induction in HepG2 cells and primary hepatocytes.

doi: 10.3945/jn.110.121277

Figure Lengend Snippet: FIGURE 5 Nrf2-dependent antioxidant capacity in HepG2 cells (A,B) or primary murine hepatocytes (C). (A) The GSH concentration was measured in lysates of cells treated with 300 mmol/L t-BHP and/ or 30 mmol/L ajoene. (B) Viability of HepG2 cells treated with 3–30 mmol/L ajoene for 12 h and continuously incubated with 300 mmol/L t-BHP for 12 h. (C) DCFH oxidation in primary hepatocytes from Nrf2 gene knockout (Nrf22/2) and wildtype (WT) mice exposed to ajoene and/or t-BHP as described above. Values are means 6 SD, n = 4. Means without a common letter differ, P , 0.05.

Article Snippet: Antibodies directed against phospho-PKCd, phospho-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), phospho-c-Jun NH2-terminal kinase (JNK) (Thr183/ Tyr185), phospho-Akt, and lamin A/C were obtained from Cell Signaling Technology, whereas that of phospho-Nrf2 (Ser40) was supplied from Novus Biologicals.

Techniques: Concentration Assay, Incubation, Gene Knockout

Journal: The Journal of nutrition

Article Title: Ajoene, a stable garlic by-product, has an antioxidant effect through Nrf2-mediated glutamate-cysteine ligase induction in HepG2 cells and primary hepatocytes.

doi: 10.3945/jn.110.121277

Figure Lengend Snippet: FIGURE 6 A schematic diagram illustrating the proposed mechanism by which ajoene protects cells against oxidative stress. Ajoene activates PKCd, which leads to Nrf2 activation. Ajoene inhibits interaction between Keap1 and Nrf2, thereby repressing Nrf2 ubiquitination, as did t-BHQ. Nrf2 activation by ajoene contributes to GCLC and GCLM induction and restoration of cellular GSH concentra- tion, which antagonizes oxidative stress.

Article Snippet: Antibodies directed against phospho-PKCd, phospho-extracellular signal-regulated kinase (ERK) (Thr202/Tyr204), phospho-c-Jun NH2-terminal kinase (JNK) (Thr183/ Tyr185), phospho-Akt, and lamin A/C were obtained from Cell Signaling Technology, whereas that of phospho-Nrf2 (Ser40) was supplied from Novus Biologicals.

Techniques: Activation Assay, Ubiquitin Proteomics

Journal: BMC biology

Article Title: Transcriptomic signatures of oxytosis/ferroptosis are enriched in Alzheimer's disease.

doi: 10.1186/s12915-025-02235-6

Figure Lengend Snippet: Fig. 5 Mechanistic validation of the TrioSig gene set in cell culture. A Diagram of the ISR and NRF2 pathways. B Levels of peIF2α, total eIF2α, NRF2, ATF4, and actin after treatment of HT22 cells with 5 mM glutamate, 500 nM erastin, and 250 nM RSL3 at various timepoints were assessed by western blotting and quantified (n = 3–4/condition). C Levels of NRF2, ATF4, and actin after knockdown with control (Ct), Nrf2, or Atf4 siRNAs (n = 7/condition). D Survival of HT22 cells exposed to varying concentrations of glutamate, erastin, or RSL3 after knockdown with Ct, Nrf2, or Atf4 siRNAs (n = 4/condition). E Levels of NRF2, ATF4, and actin in MC65 cells without (− Aβ) and with (+ Aβ) intracellular Aβ aggregation after 1 day (d1) and 2 days (d2), assessed by western blotting and quantified (n = 3/condition). F Levels of NRF2, ATF4, and actin after knockdown with control (Ct), Nrf2, or Atf4 siRNAs (n = 3/condition). G Survival of MC65 cells exposed to Aβ toxicity after knockdown with Ct, Nrf2, or Atf4 siRNAs 3 days later (n = 4–5/condition). Two-way repeated measures ANOVA and Tukey’s multiple comparisons test, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. All data are mean ± SD

Article Snippet: Twenty-four hours after seeding, cells were transfected with siRNA against Atf4 (Santa Cruz, sc-351113, 17 pmol) or Nrf2 (Santa Cruz, sc-37049, 17 pmol), using lipofectamine (RNAiMAX, ThermoFisher) as the transfection reagent and Opti-MEM (GibcoTM) as the transfection medium.

Techniques: Biomarker Discovery, Cell Culture, Western Blot, Knockdown, Control

Journal: bioRxiv

Article Title: Alterations of redox and iron metabolism accompany development of HIV latency

doi: 10.1101/549014

Figure Lengend Snippet: (A) Subcellular localization of Nrf2 in CD4 + T-cells infected with HIV-1 or mock infected (3 dpi) as analyzed by biochemical fractionation. (B) Nuclear localization (left) and content (right) of Nrf2 in HIV-1 RNA + and HIV-1 RNA - cells (7 dpi) as measured by combining IF and HIV-1 RNA FISH. Scale bar = 2μm. n= number of cells from 3 donors. (C, D) Time course of the relative (infected vs mock infected) mRNA (C) and protein (D) levels of main targets of Nrf2 during the transition from productive (3-9 dpi) to latent (14 dpi) infection as measured by qPCR and western blot, respectively. TrxR1 and HMOX-1 were probed upon membrane stripping. (E) Comparison of the effect of different HIV-1 mutations on the mean relative (infected vs mock infected) mRNA level of the Nrf2 targets described in panel D. Values shown in panel B were calculated as nuclear corrected total cell fluorescence (as in) and analyzed by two-tailed unpaired t -test. For panels C, E, raw data were first normalized using GAPDH as housekeeping control and then expressed as Log 2 fold mRNA expression in infected vs mock infected cells (calculated using the 2-ΔΔCTmethod . In panel E the average of the six genes listed in Panel C is shown. For both panels data were analyzed by-two way ANOVA followed by Turkey’s post-test for multiple comparisons. ** P <0.01; *** P <0.001; **** P <0.001. Trx= thioredoxin; NQO1= NAD(P)H Quinone Dehydrogenase 1; HMOX-1= Heme Oxygenase 1; G6PD= glucose-6-phosphate dehydrogenase; GCLC= Glutamate—cysteine ligase; TrxR1= thioredoxin reductase 1.

Article Snippet: IF for Nrf2 (mab3925, 1:250; R&D systems, Minneapolis, MN) or PML (sc/966x, 1:500; Santa Cruz Biotechnology, Dallas, TX, USA) for 1-2 hr at RT.

Techniques: Infection, Fractionation, Western Blot, Membrane, Stripping Membranes, Comparison, Fluorescence, Two Tailed Test, Control, Expressing

Journal: NPJ Science of Food

Article Title: From rosemary and coffee to bioactive nanovesicles: exploring new frontiers in food functional ingredients

doi: 10.1038/s41538-026-00723-9

Figure Lengend Snippet: Nrf2 ( A ) and COX-2 ( B ) protein levels in C2C12 cells pretreated with RVs and CVs 39 × 10 6 vesicles/mL in oxidative stress-induced damage. The data points represent the averages ±SD of three independent experiments in duplicate. All datasets were analyzed by one-way ANOVA followed by Tukey’s post hoc test. Different lowercase letters indicate a significant difference ( p 0.05) between different treatments. Control: untreated cells. RVs rosemary-derived vesicles, CVs coffee-derived vesicles. 7.8 × 10 6 vesicles/mL and 39 × 10 6 vesicles/mL correspond to 0.1 and 0.5 mg nanovesicles/mL.

Article Snippet: The antibody against Collagen Type I (Cat No. 66761-1-Ig) and Fatty Acis Synthase (FASN, Cat No. 10624-2-AP), COX2/Cyclooxygenase 2 (Cat No. 12375-1-AP) and nuclear factor erythroid 2-related factor 2 (NRF2) (Cat No. 66504-1-Ig) were from Proteintech.

Techniques: Control, Derivative Assay