rabbit anti human nrf2 primary antibody  (Proteintech)

Journal: Clinical and Translational Radiation Oncology

Article Title: Differential regulation of radioadaptation by quercetin between human normal and cancer cells

doi: 10.1016/j.ctro.2025.101099

Figure Lengend Snippet: Quercetin restores NRF2 nuclear translocation in radioadapted MCF10A cells. Representative immunofluorescence images of NRF2 (green) and DAPI (blue) and quantification of the nuclear-to-cytoplasmic NRF2 fluorescence ratio in MCF10A cells 24 h after 5 Gy irradiation with or without prior LDRT and quercetin treatment (Scale bar: 10 μm). Data is shown as mean ± SEM. * p < 0.05, ** p < 0.01.

Article Snippet: Cells were incubated with rabbit anti-human NRF2 primary antibody (1:200; Proteintech, 16396–1-AP), and detection was performed using goat anti-rabbit Alexa Fluor 488 (1:500, Invitrogen, 11001).

Techniques: Translocation Assay, Immunofluorescence, Fluorescence, Irradiation

Journal: Genes & Diseases

Article Title: TIGAR promotes osteogenic differentiation and ameliorates glucocorticoid-induced osteoporosis via autophagy-Nrf2-ROS axis

doi: 10.1016/j.gendis.2025.101735

Figure Lengend Snippet: TIGAR activated nuclear factor erythroid-2 related factor (Nrf2) to reduce dexamethasone (Dex)-induced oxidative stress through inducing autophagy. Bone marrow mesenchymal stem cells (BMSCs) were transfected with TIGAR overexpression plasmid and treated with Dex. (A, B) Immunofluorescence staining of Nrf2 of BMSCs and the quantification of the integrated optical density (IOD) per field. Scale bars, 50 μm. (C, D) Western blot analysis and quantification of Nrf2 expression in extracted nuclear proteins. (E, F) ROS level under Dex treatment in BMSCs with or without administration of 10 nM Nrf2 inhibitor (ML385) after transfecting with TIGAR overexpression plasmid, and the quantification of IOD per filed. Scale bars, 100 μm. BMSCs were treated with Dex and chloroquine (CQ) (20 μM) after transfecting with TIGAR overexpression plasmid. (G–I) Western blot analysis and quantification of p62 and LC3-II expression under different treatments. (J, K) Representative images of mCherry-GFP-LC3 puncta and number of autophagosomes (yellow) (analyzed by Pearson's correlation). Scale bars, 50 μm. (L–N) Western blot analysis and quantification of Nrf2 and kelch-associated protein 1 (Keap1) expression under different treatments. (O, P) The immunofluorescence staining of Nrf2 in BMSCs and the quantification of the IOD per field. Scale bars, 50 μm. (Q, R) Representative immunofluorescence images of LC3 and Keap1. Pearson's correlation of co-localization is shown in the bar graph format from the three independent experiments analyzed. Scale bars, 50 μm. (S, T) Representative images of ROS and the quantification of the IOD per field. Scale bars, 100 μm. Data are shown as mean ± SEM. n = 3, biologically independent samples. Two-way analysis of variance (ANOVA) with Tukey's multiple comparisons test was used to assess statistical significance. ∗ p < 0.05, ∗∗ p < 0.01. NC, negative control. OE, TIGAR overexpression plasmid.

Article Snippet: Nrf2 inhibitor (10 μM) (ML385, TargetMol Chemicals Inc., USA, CAS846557-71-9) and chloroquine (CQ, TargetMol Chemicals Inc., CAS54-05-7) in 20 μg/mL were administered to cells to explore the underlying mechanisms.

Techniques: Transfection, Over Expression, Plasmid Preparation, Immunofluorescence, Staining, Western Blot, Expressing, Negative Control

Journal: Redox Biology

Article Title: Beyond CFTR: Ivacaftor's role in restoring cellular redox balance and preventing ferroptosis

doi: 10.1016/j.redox.2025.103944

Figure Lengend Snippet: Ivacaftor reduces lipid peroxidation and protects CFBE cells from erastin-induced ferroptosis. A-B. The bar graphs show the ratio FITC-A/PE-A mean fluorescence intensity (oxidized/reduced) C11-BODIPY. Error bars are mean ± SD. A. CFBE cells manifest higher levels of lipid hydroperoxides compared to 16HBE after 24h in a 12-well plate. Paired Student's t -test (∗p < 0.05). B. CFBE cells were treated 24, 48, or 72h with elexacaftor (VX-445), tezacaftor (VX-661), lumacaftor (VX-809) or ivacaftor (VX-770) at the indicated concentrations. Vehicle DMSO was included as negative control, while RTA-408 as positive control. Data are expressed as fold change to DMSO. Ivacaftor reduces lipid hydroperoxides level in CFBE cells already after 24h of treatment. Ordinary one-way ANOVA multiple comparison vs. DMSO with Dunnett's correction (∗∗p < 0.01; ∗∗∗p < 0.001). C -D. The bar graphs show resazurin-based cell viability of CFBE cells, expressed as percentage relative to vehicle DMSO. Cells were treated with erastin (Era) alone or in combination (blue dots). Ordinary one-way ANOVA multiple comparison vs. Era with Dunnett's correction (∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001). C. VX-770 protects CFBE cells from Era-induced ferroptosis already at 24h and this effect persists over the 72h treatment. The Nrf2 activator RTA-408 does not protect from Era like vehicle DMSO, while ferroptosis inhibitor Fer-1 was included as a positive control. D. At 24h VX-770's protective ability in CFBE cells is maintained also when combined with CFTR correctors and in the triple-combination VX-770, VX-445 and VX-661 (ETI).

Article Snippet: Rabbit anti-xCT polyclonal antibody 1:1000 (26864-1-AP, Proteintech) Rabbit anti-Nrf2 polyclonal antibody 1:1000 (16396-1-AP, Proteintech) , Goat anti-rabbit IgG HRP conjugate, 1:5000 (NEF812001EA, Perkin Elmer).

Techniques: Fluorescence, Negative Control, Positive Control, Comparison

Journal: Redox Biology

Article Title: Beyond CFTR: Ivacaftor's role in restoring cellular redox balance and preventing ferroptosis

doi: 10.1016/j.redox.2025.103944

Figure Lengend Snippet: Ivacaftor modulates the Nrf2-based antioxidant response in CFBE cells. A. Representative Western blot analysis of Nrf2 protein. CFBE cells display a decreased level of Nrf2 compared to 16HBE cells. Values in the bar graph represent normalized integrated density of Western blot, expressed as fold change to 16HBE. Error bars are mean ± SD. Unpaired Student's t -test (∗∗p < 0.01). Representative Western blot of Nrf2 (B) or xCT (C) proteins in CFBE cells treated 24, 48, or 72h with ivacaftor (VX-770) 10 μM. Vehicle DMSO was used as control. Filled dots correspond to addition of the compound/vehicle. Values in the bar graphs represent normalized integrated density of Western blot and are expressed as fold change to DMSO. Error bars are mean ± SD. Unpaired Student's t -test (∗p < 0.05; ∗∗∗p < 0.001). VX-770 significantly increases Nrf2 level at 24h (B) and xCT at 48h (C) . D. 72-h treatment with VX-770 10 μM significantly increases total glutathione (tGSH) in CFBE cells. RTA-408 was included as positive control. tGSH was measured with Tietze assay and normalized to total intracellular proteins. Each point represents an independent biological replicate. Error bars are mean ± SD. Ordinary one-way ANOVA multiple comparison vs. DMSO with Dunnett's correction (∗p < 0.05; ∗∗p < 0.01). E. Similarly to the Nrf2 activator RTA-408 included as positive control, VX-770 10 μM increases the normalized relative luciferase activity in CFBE cells transiently transfected with the pGL3-8xARE-luciferase Nrf2 reporter plasmid. Each point represents an independent biological replicate and is expressed as fold change to DMSO. Error bars are mean ± SD. Ordinary one-way ANOVA multiple comparison vs. DMSO with Dunnett's correction (∗p < 0.05; ∗∗p < 0.01).

Article Snippet: Rabbit anti-xCT polyclonal antibody 1:1000 (26864-1-AP, Proteintech) Rabbit anti-Nrf2 polyclonal antibody 1:1000 (16396-1-AP, Proteintech) , Goat anti-rabbit IgG HRP conjugate, 1:5000 (NEF812001EA, Perkin Elmer).

Techniques: Western Blot, Control, Positive Control, Comparison, Luciferase, Activity Assay, Transfection, Plasmid Preparation

Journal: Redox Biology

Article Title: Beyond CFTR: Ivacaftor's role in restoring cellular redox balance and preventing ferroptosis

doi: 10.1016/j.redox.2025.103944

Figure Lengend Snippet: Ivacaftor's regulates FSP1 protein level but does not base its anti-ferroptotic activity on it. A. Representative Western blot of FSP1 protein in CFBE cells treated 24, 48, or 72h with ivacaftor (VX-770) 10 μM. Vehicle DMSO was used as control. Filled dots correspond to addition of the compound/vehicle. Values in the bar graphs represent normalized integrated density of Western blot and are expressed as fold change to DMSO. Error bars are mean ± SD. Unpaired Student's t -test (∗p < 0.05). VX-770 significantly increases FSP1 level at 24h. Western blot validation of the CRISPR-Cas9-generated CFBE Nrf2 KO (B) and FSP1 KO (C) cells, compared to the non-target control (N.T.) CFBE cell line. D. Resazurin-based cell viability assay of N.T., Nrf2 KO and FSP1 KO CFBE cells treated 24h with the ferroptosis inducer erastin. Concentrations of erastin varied in a range between 0 and 10 μM. VX-770 5 μM efficiently protected each cell line from erastin-induced ferroptosis, thus suggesting that VX-770's activity does not depend on Nrf2 and FSP1. Each point represents the mean ± SD of at least three independent biological replicates.

Article Snippet: Rabbit anti-xCT polyclonal antibody 1:1000 (26864-1-AP, Proteintech) Rabbit anti-Nrf2 polyclonal antibody 1:1000 (16396-1-AP, Proteintech) , Goat anti-rabbit IgG HRP conjugate, 1:5000 (NEF812001EA, Perkin Elmer).

Techniques: Activity Assay, Western Blot, Control, Biomarker Discovery, CRISPR, Generated, Viability Assay