Journal: Antioxidants (Basel, Switzerland)

Article Title: Photobiomodulation Controls Keratinocytes Inflammatory Response through Nrf2 and Reduces Langerhans Cells Activation.

doi: 10.3390/antiox12030766

Figure Lengend Snippet: Figure 2. PBM enhances Nrf2-pathway activation in DNCB-stimulated KCs. KCs were treated with vehicle (DMSO 0.1%) or DNCB (25 µM) and illuminated with either wavelength: 660 nm (red light) or 520 nm (green light). (A) Nrf2 accumulation was assessed by Western blot 3 h after illumination. Histograms correspond to densitometric analysis relative to controls and are adjusted to the stain-free blot. (B) ELISA-based measurement of DNA-binding activity for activated Nrf2 in the nuclear extract, 4 h after KCs treatment. (C) Nrf2 target antioxidant gene expression: HO-1, GCLC, and NQO1, were assessed by RT-qPCR, 3 h or 6 h after light treatment. Data represent mean ± SEM of at least five independent experiments. Two-Way ANOVA followed by Tukey post hoc test, * p < 0.05; ** p < 0.01; *** p < 0.001 vs. the relative untreated control at 3 h or 6 h; $ p < 0.05; $$ p < 0.01; $$$ p < 0.001 vs. DNCB at 3 h.

Article Snippet: The membrane was blocked for 1 h at room temperature in Trisbuffered saline with 5% nonfat dry milk and incubated overnight at 4 ◦C with primary rabbit anti-Nrf2 Ab (1:1000 dilution in Tris-buffered saline, 3% BSA; Proteintech®, Ref. 16396-1-AP).

Techniques: Activation Assay, Western Blot, Staining, Enzyme-linked Immunosorbent Assay, Binding Assay, Activity Assay, Gene Expression, Quantitative RT-PCR, Control

Journal: Antioxidants (Basel, Switzerland)

Article Title: Photobiomodulation Controls Keratinocytes Inflammatory Response through Nrf2 and Reduces Langerhans Cells Activation.

doi: 10.3390/antiox12030766

Figure Lengend Snippet: Figure 3. The anti-inflammatory effect of red light in KCs is Nrf2-dependent. KCs were transfected for 48 h by si-Nrf2 or si-random, and then treated with DNCB and exposed to red light. (A) Nrf2 accumulation in the knockdown model was assessed by Western blot 3 h after treatment. The blot is representative of 3 independent experiments. Histograms correspond to densitometric analysis relative to untreated si-random and are adjusted to the stain-free blot. Quantifying the expression of Nrf2 target genes encoding HO-1, NQO1, and GCLC in transfected KCs were assessed using RT-qPCR 3 h after treatment. Induction values were the ratio between gene expressions in treated cells versus gene expression in untreated si-random control. (B) Pro-inflammatory cytokines gene expression: TNF-α, IL-6 and IL-8, were assessed by RT-qPCR, 3 h after light treatment. Results are expressed as fold change compared to the untreated si-random control. (C) The inflammatory cytokines level was determined by Meso Scaled Discovery technology in the supernatant of KCs after 6 h of exposure to DNCB with or without red light illumination. Reported data are mean ± SEM of 6 independent experiments. Two-Way ANOVA followed by Tukey post hoc test, * p < 0.05; ** p < 0.01; *** p < 0.001 vs. untreated si-random; $ p < 0.05; $$ p < 0.01; $$$ p < 0.001 vs. DNCB-treated si-random, $$′ p < 0.01; $$$′ p < 0.001 vs. DNCB + 660 nm-treated si-random; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. untreated si-Nrf2. ns means non-significant different.

Article Snippet: The membrane was blocked for 1 h at room temperature in Trisbuffered saline with 5% nonfat dry milk and incubated overnight at 4 ◦C with primary rabbit anti-Nrf2 Ab (1:1000 dilution in Tris-buffered saline, 3% BSA; Proteintech®, Ref. 16396-1-AP).

Techniques: Transfection, Knockdown, Western Blot, Staining, Expressing, Quantitative RT-PCR, Gene Expression, Control

Journal: BioMed Research International

Article Title: Different Parts of the Chicken Embryo Egg Improve D-Galactose-Induced Aging in a Mice Model

doi: 10.1155/2021/6654683

Figure Lengend Snippet: Immunohistochemistry staining for HO-1 and NRF-2 in liver sections. (a) Young, (b) D-gal, (c) D-gal/CEr, (d) D-gal/CEw, (e) D-gal/CEp, (f) HO-1 positive cells counting, and (g) NRF-2 positive cells counting. Magnification ×200. ∗ P < 0.05 or ∗∗ P < 0.01 compared with the D-gal group.

Article Snippet: Tissue sections were heated in 10 mmol/L sodium citrate (pH 6.0) in a water bath at 95°C for 10 min to expose the tissue's antigens and then incubated at 4°C overnight with diluted (1 : 200) mouse monoclonal anti-heme oxygenase-1 (HO-1) antibody or rabbit polyclonal antinuclear factor erythroid-2-related factor 2 (NRF-2) antibody (Novus Biologicals LLC, Centennial, CO).

Techniques: Immunohistochemistry, Staining

Journal: Molecular medicine reports

Article Title: Protective effect of Porphyra yezoensis glycoprotein on D-galactosamine‑induced cytotoxicity in Hepa 1c1c7 cells.

doi: 10.3892/mmr.2015.3244

Figure Lengend Snippet: Figure 5. Effect of PYGP on D‑GaIN‑induced expression of Nrf2, Nqo1, GST and HO‑1 in Hepa 1c1c7 cells. Cells were pre‑treated with PYGP (20 and 40 µg/ml) for 24 h and then administered 20 mM D‑GaIN with PYGP (20 and 40 µg/ml) for 24 h. Cell pellets were then collected using lysis buffer and western blot analysis was performed in order to determine protein expression levels of Nrf2, Nqol, GST and HO‑1. PYGP, Porphyra yezoensis glycopro tein; D‑GaIN, D‑galactosamine; Nrf2, nuclear factor erythroid 2‑related factor 2; Nqol, quinine oxidoreductase 1; GST, glutathione s‑transferase; HO‑1, heme oxygenase 1.

Article Snippet: The transferred membrane was blocked at room temperature with 1% bovine serum albumin (BSA) in Tris‐buffered saline with Tween 20 [TBS-T; 10 mM Tris-HCl (pH 7.5), 150 mM NaCl and 0.1% Tween 20] and then incubated, with agitation, with the indicated primary antibodies: Rabbit anti-mouse ERK immunoglobulin G (IgG) polyclonal antibody [diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at room temperature (RT)], rabbit anti-phosphorylated (p)-ERK IgG polyclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), mouse anti-mouse JNK IgG monoclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), mouse anti-mouse p-JNK IgG monoclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), rabbit anti-mouse p38 IgG polyclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), mouse anti-mouse p-p38 IgG monoclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), rabbit anti-mouse Nrf2 IgG polyclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), goat anti-mouse Nqo1 IgG polyclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), mouse anti-mouse GST IgG monoclonal antibody (diluted 1:1,000 with BSA/TBS-T; incubated for 4 h at RT), and rabbit anti-mouse heme oxygenase (HO)-1 IgG polyclonal antibody (diluted 1:1,000 with BSA/ TBS-T; incubated for 4 h at RT), which were all purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Expressing, Lysis, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Synergistic Effects of Vitis vinifera L. and Centella asiatica against CCl 4 -Induced Liver Injury in Mice

doi: 10.3390/ijms241411255

Figure Lengend Snippet: List of specific primary and secondary antibodies used for immunoblotting.

Article Snippet: 6 , Nrf-2 , 5% BSA , 1:1000 , Rabbit IgG , Gene Tex , GTX103322.

Techniques: Western Blot, Blocking Assay

Journal: European Journal of Medical Research

Article Title: Quercetin alleviates radiation-induced erectile dysfunction by modulating oxidative stress and apoptosis through the Nrf2/HO-1 pathway

doi: 10.1186/s40001-026-03978-w

Figure Lengend Snippet: In vitro validation of the protective effects of quercetin against radiation-induced injury in endothelial cells. A , B Cell viability of endothelial cells at 24 h and 48 h after exposure to increasing radiation doses (0–10 Gy), assessed by CCK-8 assay and expressed as a percentage of the non-irradiated control. C , D Cell viability at 24 h and 48 h in non-irradiated cells treated with increasing concentrations of quercetin, showing the range of doses with minimal cytotoxicity. E Effects of radiation and selected quercetin concentrations on endothelial cell viability at 48 h (Control, IR, and IR + Que at the indicated doses). F Representative immunofluorescence images of intracellular ROS (red) and nuclei (DAPI, blue) in the Control, Que, IR and IR + Que groups (scale bar = 50 μm). G Quantification of ROS fluorescence intensity (fold of Control) showing radiation-induced ROS accumulation and its attenuation by quercetin. H Representative immunofluorescence images of Nrf2 (red) and nuclei (DAPI, blue) in each group (scale bar = 50 μm). I Quantification of Nrf2 fluorescence intensity (fold of Control). J Representative immunofluorescence images of HO-1 (green) and nuclei (DAPI, blue) in each group (scale bar = 50 μm). K Quantification of HO-1 fluorescence intensity (fold of Control). Data are presented as mean ± SD and were analysed by one-way ANOVA followed by Tukey’s post hoc test (n = 3 independent experiments). (* p < 0.05 compared with Control group; ** p < 0.01 compared with Control group; ## p < 0.01 compared with IR group)

Article Snippet: After fixation, cells were permeabilised with 0.3% Triton X-100 in PBS for 10 min at room temperature, followed by blocking with 5% BSA for one h. After blocking, cells were incubated overnight at 4 °C with primary antibodies diluted in BSA blocking buffer: anti-Nrf2 antibody (Proteintech, 80593-1-RR, 1:250) or anti-HO-1 antibody (Proteintech, 66743-1-Ig, 1:250).

Techniques: In Vitro, Biomarker Discovery, CCK-8 Assay, Irradiation, Control, Immunofluorescence, Fluorescence

Journal: European Journal of Medical Research

Article Title: Quercetin alleviates radiation-induced erectile dysfunction by modulating oxidative stress and apoptosis through the Nrf2/HO-1 pathway

doi: 10.1186/s40001-026-03978-w

Figure Lengend Snippet: Expression and localisation analysis of Nrf2/HO-1 in the penile tissues of different rat groups. A Immunofluorescence staining images showing the expression of Nrf2 (red) in the control, model, and treatment groups (Que-L and Que-H). DAPI staining (blue) marks the cell nuclei, and merged images show the localisation of Nrf2 relative to the nuclei (DAPI). Scale bar = 20 μm. B Quantification of Nrf2-positive areas, showing a significant decrease in the model group compared to the control group, and partial recovery in the Que-L and Que-H groups. C Immunofluorescence staining images showing the expression of HO-1 (green) in the control, model, and treatment groups (Que-L and Que-H). DAPI staining (blue) marks the cell nuclei, and merged images show the localisation of HO-1 relative to the nuclei (DAPI). Scale bar = 20 μm. D Quantification of HO-1-positive areas, showing a significant decrease in the model group compared to the control group, and partial recovery in the Que-L and Que-H groups. E Western blot analysis was used to detect the expression levels of Nrf2 and HO-1 in penile tissues, with β-actin serving as the loading control. F , G Quantification of Nrf2 and HO-1 protein expression levels normalised to β-actin. Data are presented as mean ± SD and were analysed by one-way ANOVA followed by Tukey’s post hoc test (n = 8 per group). (* p < 0.05 compared with Control group; ** p < 0.01 compared with Control group; # p < 0.05 compared with Model group; ## p < 0.01 compared with Model group; & p < 0.05 compared with Que-L group)

Article Snippet: After fixation, cells were permeabilised with 0.3% Triton X-100 in PBS for 10 min at room temperature, followed by blocking with 5% BSA for one h. After blocking, cells were incubated overnight at 4 °C with primary antibodies diluted in BSA blocking buffer: anti-Nrf2 antibody (Proteintech, 80593-1-RR, 1:250) or anti-HO-1 antibody (Proteintech, 66743-1-Ig, 1:250).

Techniques: Expressing, Immunofluorescence, Staining, Control, Western Blot

Journal: European Journal of Medical Research

Article Title: Quercetin alleviates radiation-induced erectile dysfunction by modulating oxidative stress and apoptosis through the Nrf2/HO-1 pathway

doi: 10.1186/s40001-026-03978-w

Figure Lengend Snippet: Mechanistic scheme showing how quercetin alleviates radiation-induced erectile dysfunction by modulating oxidative stress and apoptosis via activation of the Nrf2/HO-1 pathway

Article Snippet: After fixation, cells were permeabilised with 0.3% Triton X-100 in PBS for 10 min at room temperature, followed by blocking with 5% BSA for one h. After blocking, cells were incubated overnight at 4 °C with primary antibodies diluted in BSA blocking buffer: anti-Nrf2 antibody (Proteintech, 80593-1-RR, 1:250) or anti-HO-1 antibody (Proteintech, 66743-1-Ig, 1:250).

Techniques: Activation Assay