primary antibodies against nrf2  (Cell Signaling Technology Inc)

Journal: Frontiers in Pharmacology

Article Title: Coreopsis tinctoria Nutt. attenuates ultraviolet A photodamage by suppressing endoplasmic reticulum stress-induced apoptosis via Nrf2 crosstalk

doi: 10.3389/fphar.2025.1686234

Figure Lengend Snippet: CT alleviates UVA-induced ER stress and apoptosis via Nrf2 in HaCaT cells. (A) HaCaT cells were subjected to UVA radiation and treated with 50 or 100 μg/mL CT and/or siNrf2. Western blot assays were performed to detect the levels of ATF6, phosphorylated elF2a (p-elF2a), CHOP, GADD34, Cleaved caspase9, Cleaved caspase3, JNK, phosphorylated JNK (p-JNK), Bax, BCL2, cytoplasmic and nuclear NRF2 and HO-1. (B) Flow cytometry analysis was performed to determine cell apoptosis. Quantification of the percentage of apoptotic cells in different treatments. (C) Heatmap generated the data from RNAseq. Genes are all related to NRF2 downstream anti-oxidant or detoxification target genes. The figure was generated by SRplot . N = 4. * p ≤ 0.05; ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: After incubation with an endogenous peroxidase blocker (ZSGB-BIO, PV-6000), the slides were blocked with 3% BSA for 1 h. Primary antibodies against NRF2 (Proteintech, 16396-1-AP), Caspase12 (Proteintech, 55238-1-AP), and CHOP (Proteintech, 15204-1-AP) were incubated overnight at 4 °C at a dilution of 1:200 each.

Techniques: Western Blot, Flow Cytometry, Generated

Journal: Frontiers in Pharmacology

Article Title: Coreopsis tinctoria Nutt. attenuates ultraviolet A photodamage by suppressing endoplasmic reticulum stress-induced apoptosis via Nrf2 crosstalk

doi: 10.3389/fphar.2025.1686234

Figure Lengend Snippet: Therapeutic effects of CT on macroscopic and histological changes induced by ER stress in UVA-photodamaged mouse skin. (A) Schematic representation of the experimental design for mice study. (B) Representative pictures of untreated or treated with UVA with CT or not. (C) Representative pictures of H&E and Masson staining of the skin of mice from different treatment groups on day 14. (D–F) Representative images of CHOP, NRF2, and caspase-12 for IHC analysis performed on skin sections of mice from different treatment groups on day 14.

Article Snippet: After incubation with an endogenous peroxidase blocker (ZSGB-BIO, PV-6000), the slides were blocked with 3% BSA for 1 h. Primary antibodies against NRF2 (Proteintech, 16396-1-AP), Caspase12 (Proteintech, 55238-1-AP), and CHOP (Proteintech, 15204-1-AP) were incubated overnight at 4 °C at a dilution of 1:200 each.

Techniques: Staining

Journal: Frontiers in Cell and Developmental Biology

Article Title: Perillaldehyde synergizes with ferroptosis inducers to promote ferroptotic cell death in gastric cancer

doi: 10.3389/fcell.2025.1598520

Figure Lengend Snippet: PAH induces oxidative stress while activating the P62-Keap1-Nrf2 pathway (A) AGS cells were treated with 200 μmol/L PAH for 24 h, and RNA sequencing volcano plot showed significantly differentially expressed genes. Differential genes are screened according to the threshold set by | log2(FoldChange) | and p -adj (<0.05). Blue indicates genes with downregulated expression, and red indicates genes with upregulated expression. (B) Inter-sample clustering heatmap for RNA sequencing. Green indicates low gene expression, red indicates high gene expression, and the connecting lines represent clustering results. (C) KEGG functional enrichment analysis of AGS cells treated with PAH, the results with p .adjust <0.05 and the top 20 gene numbers are selected for display. The size of the dots indicates the number of differential genes enriched in this pathway, the redder the dot color is, the more significant it is. (D) Compared with the control group, the results of Reactome pathway enrichment analysis of differential genes in AGS cells treated with PAH. The redder the dot color, the more significant it is. (E) AGS and HGC27 cells were treated with different concentrations of PAH for 24 h. Measurement of Nrf2 protein nuclear localization by laser confocal microscopy. Scale bar: 50 μm. (F,G) HGC27 and AGS cells were treated with corresponding concentration gradient PAH for 24 h. Western blotting was used to detect the protein expression of P62, Keap1, Nrf2, HMOX1 and NQO1 ( n = 3) and ImageJ software was used to quantify the protein band (Data are expressed as mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, significantly different from the control group).

Article Snippet: The cells were then incubated overnight with a primary antibody against Nrf2 (R1312-8; HUABIO), followed by incubation with a fluorescent secondary antibody (E-AB-1005; Elabscience).

Techniques: RNA Sequencing, Expressing, Gene Expression, Functional Assay, Control, Confocal Microscopy, Concentration Assay, Western Blot, Software

Journal: Frontiers in Cell and Developmental Biology

Article Title: Perillaldehyde synergizes with ferroptosis inducers to promote ferroptotic cell death in gastric cancer

doi: 10.3389/fcell.2025.1598520

Figure Lengend Snippet: PAH regulates ferroptosis through the P62-Keap1-Nrf2 pathway. (A,B) Cell viability was assessed by MTT assay after 24-h treatment with PAH and (or) siRNA Nrf2 in HGC27 and AGS cells ( n = 4). (C) Western blot assay of P62, Keap1, Nrf2, HMOX1 and NQO1 protein expression in HGC27 and AGS cells after 24 h of PAH and Fer-1 co-treatment ( n = 3). (D) Western blot assay of P62, Keap1, Nrf2, HMOX1 and NQO1 protein expression in HGC27 and AGS cells after 24 h of PAH and RSL3 co-treatment ( n = 3). (E) ImageJ software was used to quantify the protein bands in HGC27 and AGS cells after 24 h of PAH and Fer-1 co-treatment ( n = 3). (F) ImageJ software was used to quantify the protein bands in HGC27 and AGS cells after 24 h of PAH and RSL3 co-treatment ( n = 3) (Data are expressed as mean ± SD; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, significantly different from the Control or PAH group).

Article Snippet: The cells were then incubated overnight with a primary antibody against Nrf2 (R1312-8; HUABIO), followed by incubation with a fluorescent secondary antibody (E-AB-1005; Elabscience).

Techniques: MTT Assay, Western Blot, Expressing, Software, Control

Journal: World Journal of Gastroenterology

Article Title: Rhapontin activates nuclear factor erythroid 2-related factor 2 to ameliorate 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced gastrointestinal dysfunction in Parkinson's disease mice

doi: 10.3748/wjg.v31.i15.104875

Figure Lengend Snippet: Rhapontin activates nuclear factor erythroid 2-related factor 2 in the colon. A: Western Blot image and analysis of nuclear factor erythroid 2-related factor 2 (NRF2) in colon; B: Western blot image and analysis of NRF2 in substantia nigra; C: Western blot image and analysis of NRF2 in striatum. n = 10. a P < 0.05; b P < 0.01; NS: Not significant; NRF2: Nuclear factor erythroid 2-related factor 2; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; M: MPTP; R: Rhapontin; ML: ML385.

Article Snippet: Equal amounts of protein were resolved on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels, transferred onto polyvinylidene fluoride membranes, and incubated with primary antibodies against NRF2 (1:1000, Servicebio) and β-actin (1:5000, Servicebio) overnight at 4 °C.

Techniques: Western Blot