nrf2 d9j1b  (Cell Signaling Technology Inc)

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: Esophageal dysplasia in Keap1-cKO mice was abolished by Nrf2 deletion. ( A ) Immunohistochemical staining for KEAP1, NRF2, and NQO1 at 1 week after Tam administration. In KEAP1-deleted cells, NRF2 accumulated in the nucleus, and NQO1 was expressed at high levels. Scale bars: 50 μm. ( B ) NQO1 immunohistochemistry and HE staining in Keap1 floxB/floxB :: Nrf2 –/– and K5CreERT2 :: Keap1 floxB/floxB :: Nrf2 –/– mouse esophagi. Scale bars: 100 μm.

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Immunohistochemical staining, Staining, Immunohistochemistry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: NRF2 activation decreases 4 weeks after Tam treatment. ( A ) Immunohistochemical staining for NQO1. Population of NQO1-positive cells decreased over time in groups treated with each dose. Scale bars: 400 μm. ( B ) Quantification of NQO1-positive cells in the esophageal epithelium of mice treated with the 3 doses. Percentage of NQO1-positive cells was calculated as the NQO1-positive area within the total esophageal epithelial area in each mouse (n = 7–12 mice per group). ( C ) Positive correlations of percentage of NQO1-positive cells with thicknesses of keratinous and epithelial layers 1 week after administration of 3 doses of Tam (n = 29 mice). Line shows the two-tailed Pearson correlation. ( D ) Nqo1 and Gclc mRNA expression levels. Gene expression was decreased throughout the 4-week period in each group (n = 4–6 mice per group). Data are presented as means ± standard deviations. ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001 according to Welch t test.

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Activation Assay, Immunohistochemical staining, Staining, Two Tailed Test, Expressing, Gene Expression

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: NRF2-activated cells are selectively eliminated and induce DNA damage in neighboring KEAP1-normal cells. ( A ) NQO1-positive floating cells were present in the Keap1-cKO mouse esophagus. ( B ) Immunofluorescence staining for NQO1 and COL17A1. White arrows indicate COL17A1-weak areas. NRF2-activated cells exhibited weak COL17A1 staining. ( C ) COL17A1-weak areas were counted in each section of esophageal epithelium from Keap1-cKO mice (n = 9). COL17A1-weak areas were particularly abundant in NQO1-positive basal cells. ( D ) Immunofluorescence staining for NQO1 and Loricrin. ( E ) Schematic of the esophageal epithelium of Keap1-cKO mice. NQO1-positive cells exhibited weaker COL17A1 staining than KEAP1-normal cells. Scale bars: 50 μm. Data are presented as means ± standard deviations. ∗∗ P < .01, as compared using Welch t test.

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Immunofluorescence, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: NRF2-activated cells are selectively eliminated and induce DNA damage in neighboring KEAP1-normal cells. ( A ) Immunofluorescence staining for NQO1 and Ki67. The number of Ki67-positive cells was increased, especially among NRF2-activated cells, in the esophageal epithelium of Keap1-cKO mice. ( B ) Percentages of Ki67-positive cells among NQO1-negative and NQO1-positive cells in the Keap1-cKO mouse esophagus (n = 9). ( C ) The number of Ki67-positive cells in the basal layer was normalized to the length of the basement membrane (n = 12 control mice and n = 9 Keap1-cKO mice). ( D ) Ki67 mRNA expression level (n = 9 control mice and n = 10 Keap1-cKO mice). ( E ) Immunofluorescence staining for NQO1 and γH2A.X. The number of γH2A.X-positive cells was significantly increased, especially among NRF2-activated cells, in the Keap1-cKO mouse esophagus. ( F ) Percentages of γH2A.X-positive cells among NQO1-negative and NQO1-positive cells in the Keap1-cKO mouse esophagus. ( G ) Immunohistochemical staining for γH2A.X. ( H ) The number of γH2A.X-positive cells in the basal layer was normalized to the length of the basement membrane (n = 4–11 mice per group). Scale bars: 50 μm. Data are presented as means ± standard deviations. ∗ P < .05 and ∗∗∗ P < .001, as compared using Welch t test.

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Immunofluorescence, Staining, Membrane, Control, Expressing, Immunohistochemical staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: Additional NRF2 activation in Keap1-cKO::Nrf2 SA mice promotes elimination of NRF2-activated cells. ( A ) Immunohistochemical staining for NQO1. NQO1-positive cells were distributed segmentally in the esophageal epithelium of both Keap1-cKO::Nrf2 SA mice and Keap1-cKO mice. Scale bars: 400 μm. ( B ) Quantification of esophageal epithelial length. The epithelial length was greatest in Keap1-cKO::Nrf2 SA mice (n = 9–12 mice per group). ( C ) Quantification of NQO1-positive cells in the esophageal epithelium. The number of NQO1-positive cells decreased earlier in Keap1-cKO::Nrf2 SA mouse esophagus than in Keap1-cKO mouse esophagus (n = 8–10 mice per group). ( D and E ) Immunohistochemical staining for Ki67. A greater increase in number of Ki67-positive cells was observed in the Keap1-cKO::Nrf2 SA mouse esophagus than in the Keap1-cKO mouse esophagus. Scale bars: 50 μm. ( F and G ) Immunohistochemical staining for γH2A.X. Greater increase in number of γH2A.X-positive cells was observed in Keap1-cKO::Nrf2 SA mouse esophagus than in Keap1-cKO mouse esophagus. Scale bars: 50 μm. Data are presented as means ± standard deviations. ∗ P < .05, ∗∗ P < .01, and ∗∗∗ P < .001 according to Welch t test.

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Activation Assay, Immunohistochemical staining, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: Accelerated proliferation of KEAP1-normal cells. ( A ) UMAP visualization of cells isolated from control and Keap1-cKO mouse esophagi. Left panel , 4853 cells are colored on basis of cell type. Right panel , 2472 and 2381 cells are colored on basis of their origin from control and Keap1-cKO mice, respectively. ( B ) Nqo1 expression in epithelial cells. ( C ) GSEA enrichment plot comparing Nqo1 Low and Nqo1 High epithelial cells using the NRF2 pathway gene set obtained from Wiki Pathways. ( D ) GSEA enrichment plot comparing Nqo1 Low and Nqo1 High epithelial cells using hallmark gene sets. Left panel shows the gene sets enriched in Nqo1 High cells. Only the reactive oxygen species pathway was significantly enriched among the hallmark gene sets. The right two panels show the hallmark gene sets enriched in Nqo1 Low cells. ( E ) GSEA enrichment plot comparing control and Nqo1 Low epithelial cells. The gene sets E2F targets and G2/M checkpoint were significantly enriched in Nqo1 Low cells.

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Isolation, Control, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Selective Elimination of NRF2-Activated Cells by Competition With Neighboring Cells in the Esophageal Epithelium

doi: 10.1016/j.jcmgh.2022.09.004

Figure Lengend Snippet: Antibodies Used for Immunohistochemistry and Immunofluorescence

Article Snippet: NRF2 (D9J1B) , Rat monoclonal , Cell Signaling Technology , 14596 , 1:400 , —.

Techniques: Immunohistochemistry