protein 1 keap1  (Cell Signaling Technology Inc)

Journal: Molecular Medicine Reports

Article Title: Pathological mechanism of ferroptosis in a rat model of α-naphthyl isothiocyanate-induced chronic cholestasis

doi: 10.3892/mmr.2026.13802

Figure Lengend Snippet: Abnormal expression of antioxidant and iron metabolism-related proteins in liver samples of rats with chronic cholestasis. Control group was fed a chow diet, whereas the ANIT and DFO-treated groups were intragastrically administered ANIT olive oil solution with or without DFO (n=8/group). (A) Changes in ferroptosis antioxidant-related protein expression. The blots shown are representative of three independent experiments. A dotted line indicates that the lanes were non-adjacent on the original gel. All target protein bands and their corresponding loading control bands shown side-by-side were derived from the same membrane. Relative expression levels of (B) Nrf2, (C) Keap1, (D) XCT, (E) HO-1 and (F) GPX4. (G) Changes in iron metabolism-related protein expression. The blots shown are representative of three independent experiments. A dotted line indicates that the lanes were non-adjacent on the original gel. All target protein bands and their corresponding loading control bands shown side-by-side were derived from the same membrane. Relative expression levels of (H) TFR1, (I) FPN1, (J) DMT1, (K) Steap3 and (L) FTH1. Data are presented as the mean ± SD. P-values were determined by one-way ANOVA. *P<0.05, **P<0.01, ***P<0.001. ANIT, α-naphthyl isothiocyanate; DFO, deferoxamine; Nrf2, nuclear factor erythroid-2-related factor 2; Keap1, Kelch-like ECH-associated protein 1, XCT, cystine/glutamate transporter; HO-1, heme oxygenase 1; GPX4, glutathione peroxidase 4; TFR1, transferrin receptor 1; FPN1, ferroportin 1; DMT1, divalent metal transporter 1; Steap3, six-transmembrane epithelial antigen of the prostate 3; FTH1, ferritin heavy chain 1.

Article Snippet: The Kelch-like ECH-associated protein 1 (Keap1) antibody (cat. no. bs-3648R) was purchased from BIOSS.

Techniques: Expressing, Control, Derivative Assay, Membrane

Journal: Molecular Medicine Reports

Article Title: Pathological mechanism of ferroptosis in a rat model of α-naphthyl isothiocyanate-induced chronic cholestasis

doi: 10.3892/mmr.2026.13802

Figure Lengend Snippet: Diagram of the mechanism of ferroptosis. ASCL4, acyl-CoA synthetase long-chain family member 4; COX2, cyclooxygenase 2; DMT1, divalent metal transporter 1; FPN, ferroportin; GSSG, oxidized glutathione; GPX4, glutathione peroxidase 4; GSH, glutathione; HO-1, heme oxygenase 1; Keap1, Kelch-like ECH-associated protein 1; LIP, labile iron pool; LPCAT3, lysophosphatidylcholine acyltransferase 3; NCOA4, nuclear receptor coactivator 4; NOX1, nicotinamide adenine dinucleotide phosphate oxidase 1; NQO1, NAD(P)H quinone dehydrogenase 1; Nrf2, nuclear factor erythroid-2-related factor 2; PL-PUFA-OOH, phospholipid-polyunsaturated fatty acid hydroperoxide; SLC7A11, solute carrier family 7 member 11; Steap3, six-transmembrane epithelial antigen of the prostate 3; TFR1, transferrin receptor 1.

Article Snippet: The Kelch-like ECH-associated protein 1 (Keap1) antibody (cat. no. bs-3648R) was purchased from BIOSS.

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Hydrogen improves the efficacy of tetrandrine in the treatment of silicosis by inhibiting vascular endothelial mesenchymal transition caused by oxidative stress

doi: 10.3389/fbioe.2025.1668524

Figure Lengend Snippet: Tetrandrine concentration decreased in lung tissue and oxidative stress excessive activated in silicosis. (A,B) , HE staining and quantitative analysis of the degree of vascular stenosis of each group of mice (n = 3), scale bar, 20 μm; (C,D) , Masson staining and collagen volume fraction of each group of mice (n = 3), scale bar, 20 μm; (E,F) , Immunofluorescence and semi-quantitative of the expression of the endothelial to mesenchymal transition (EndMT)-related proteins platelet endothelial cell adhesion molecule-1 (CD31) and alfa-smooth muscle actin (α–SMA) in the lung tissues of different mouse groups (n = 3). Scale bar, 10 μm and 2 μm, Pearson’s R is the correlation coefficient—a statistical measure that indicates both the strength and direction of a linear relationship between two continuous variables, ranging from −1 (strong negative) to +1 (strong positive); (G) , The concentration of tetrandrine in lung tissue was detected by LC/MS-MS (n = 3); (H,I) , Changes of ROS levels in the lung tissues (n = 3). Scale bar, 20 μm, ROS: reactive oxygen species, DAPI: 4′,6-Diamidino-2-phenylindole dihydrochloride, Merge: ROS combines with the cell nucleus; (J) , Western blotting of Keap1 and Nrf2 expression in the lung tissues (n = 3); (K,L) , Semi-quantitative of the expression of Keap1 and Nrf2 in the lung tissues (n = 3). a P < 0.05; aa P < 0.01 vs. the control group. Data are expressed as mean ± SD.

Article Snippet: The membranes were incubated with the primary antibodies, namely, Pink1 (1:1000 dilution, Proteintech, United States, 23274-1-AP), LC3 (1:1000 dilution, Cell Signaling Technology, United States, 12741T), sequestosome 1 (SQSTM1/p62) (1:1000 dilution, Cell Signaling Technology, United States, 5114T), Parkin (1:2000 dilution, Proteintech, United States, 14060-1-AP), Kelch-like ECH-associated protein 1 (Keap1) (1:1000 dilution, Proteintech, United States, 60027-1-Ig), nuclear factor E2-related factor 2 (Nrf2) (1:2000 dilution, Proteintech, United States, 16396-1-AP), B- cell lymphoma - 2 (Bcl-2) (1:2000 dilution, Proteintech, United States, 26593-1-AP), Bcl - 2-associated X protein (Bax) (1:5000 dilution, Proteintech, United States, 50599-2-Ig), Cytochrome C (Cyto C) (1:5000, Proteintech, United States, 10993-1-AP), Caspase 3 (1:1000 dilution, Proteintech, United States, 19677-1-AP) and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:500000 dilution, Proteintech, United States, 60004-1-Ig).

Techniques: Concentration Assay, Staining, Immunofluorescence, Expressing, Liquid Chromatography with Mass Spectroscopy, Western Blot, Control

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Hydrogen improves the efficacy of tetrandrine in the treatment of silicosis by inhibiting vascular endothelial mesenchymal transition caused by oxidative stress

doi: 10.3389/fbioe.2025.1668524

Figure Lengend Snippet: Inhibiting oxidative stress can improve the concentration of tetrandrine in lung tissue in mice with silicosis. (A,B) , Changes in the ROS levels in the lung tissues (n = 3). Scale bar, 20 μm, ROS: reactive oxygen species, DAPI: 4′,6-Diamidino-2-phenylindole dihydrochloride, Merge: ROS combines with the cell nucleus; (C) , Western blotting of Keap1, Nrf2, Pink1, Parkin, Bcl-2 and Bax expression in the lung tissues (n = 3); (D,E) , Semi-quantitative of the expression of Keap1 and Nrf2 in the lung tissues of different mouse groups (n = 3); (F) , Mitochondrial injury observed under transmission electron microscopy (TEM) (n = 3). Scale bar, 2.0 (×4.0k) and 1.0 (×8.0k) μm; (G–J) , Semi-quantitative of the expression of Pink1, Parkin, Bcl-2 and Bax in the lung tissues of different mouse groups (n = 3); (K,L) , Immunofluorescence and semi-quantitative of the expression of the endothelial to mesenchymal transition (EndMT)-related proteins platelet endothelial cell adhesion molecule-1 (CD31) and alfa-smooth muscle actin (α–SMA) in the lung tissues of different mouse groups (n = 3). Scale bar, 10 μm and 2 μm; (M) , The concentration of tetrandrine in lung tissue was detected by LC/MS-MS (n = 3 biologically independent experiments). a P < 0.05; aa P < 0.01; aaa P < 0.001 vs. the control group. b P < 0.05; bb P < 0.01 vs. the model group. Data are expressed as mean ± SD.

Article Snippet: The membranes were incubated with the primary antibodies, namely, Pink1 (1:1000 dilution, Proteintech, United States, 23274-1-AP), LC3 (1:1000 dilution, Cell Signaling Technology, United States, 12741T), sequestosome 1 (SQSTM1/p62) (1:1000 dilution, Cell Signaling Technology, United States, 5114T), Parkin (1:2000 dilution, Proteintech, United States, 14060-1-AP), Kelch-like ECH-associated protein 1 (Keap1) (1:1000 dilution, Proteintech, United States, 60027-1-Ig), nuclear factor E2-related factor 2 (Nrf2) (1:2000 dilution, Proteintech, United States, 16396-1-AP), B- cell lymphoma - 2 (Bcl-2) (1:2000 dilution, Proteintech, United States, 26593-1-AP), Bcl - 2-associated X protein (Bax) (1:5000 dilution, Proteintech, United States, 50599-2-Ig), Cytochrome C (Cyto C) (1:5000, Proteintech, United States, 10993-1-AP), Caspase 3 (1:1000 dilution, Proteintech, United States, 19677-1-AP) and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:500000 dilution, Proteintech, United States, 60004-1-Ig).

Techniques: Concentration Assay, Western Blot, Expressing, Transmission Assay, Electron Microscopy, Immunofluorescence, Liquid Chromatography with Mass Spectroscopy, Control