Journal: Scientific Reports

Article Title: The cytoprotective role of DJ-1 and p45 NFE2 against human primary alveolar type II cell injury and emphysema

doi: 10.1038/s41598-018-21790-3

Figure Lengend Snippet: MMP9, CD147, cathepsin B, p18 NFE2, p45 NFE2 and ADAMTSL-4 expression in ATII cells and lung tissue obtained from non-smokers (NS), smokers (SM) and patients with emphysema (E). MMP9, CD147 and cathepsin B expression in freshly isolated ATII cells ( A ) and lung tissue ( B ) as detected by Western blotting analysis. ( C ) MMP9, CD147, cathepsin B, p45 NFE2 and ADAMTSL-4 mRNA levels in lung tissue by RT-PCR. ( D,E ) p18 NFE2, p45 NFE2 and ADAMTSL-4 expression in freshly isolated ATII cells ( D ) and lung tissue ( E ). Protein levels were analyzed by Western blotting. Densitometric analysis is also shown. *Statistically significant difference ( p < 0.05) is shown for comparison between non-smokers and smokers, between smokers and emphysema patients and between non-smokers and emphysema patients. Data are shown as the mean (±s.e.m.).

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): MMP9, CD147, ADAMTSL-4, p45 NFE2, p18 NFE2, DJ-1, phosphoserine, phosphothreonine and phosphotyrosine.

Techniques: Expressing, Isolation, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: The cytoprotective role of DJ-1 and p45 NFE2 against human primary alveolar type II cell injury and emphysema

doi: 10.1038/s41598-018-21790-3

Figure Lengend Snippet: p45 NFE2 and p18 NFE2 tyrosine phosphorylation in human ATII cells and lung tissue obtained from control non-smokers (NS) and smokers (SM) and patients with emphysema (E). Immunoprecipitation was performed in freshly isolated ATII cells ( A ) and lung tissue ( B ) followed by Western blotting analysis. Densitometric quantification is also shown. *Statistically significant difference ( p < 0.05). Data are shown as the mean (±s.e.m.).

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): MMP9, CD147, ADAMTSL-4, p45 NFE2, p18 NFE2, DJ-1, phosphoserine, phosphothreonine and phosphotyrosine.

Techniques: Immunoprecipitation, Isolation, Western Blot

Journal: Scientific Reports

Article Title: The cytoprotective role of DJ-1 and p45 NFE2 against human primary alveolar type II cell injury and emphysema

doi: 10.1038/s41598-018-21790-3

Figure Lengend Snippet: NFE2 knockdown increases cell death induced by CSE in A549 cells in vitro . ( A ) A549 cells were transfected with 100 nM NFE2 siRNA or non-targeting (NT) siRNA followed by exposure to CSE for 24 h. Representative flow cytometry images using Annexin V and PI staining are shown. ( B ) NFE2 knockdown significantly increased cell death after treatment with CSE compared to control. *Statistically significant difference ( p < 0.05). Data are shown as the mean (±s.e.m.).

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): MMP9, CD147, ADAMTSL-4, p45 NFE2, p18 NFE2, DJ-1, phosphoserine, phosphothreonine and phosphotyrosine.

Techniques: In Vitro, Transfection, Flow Cytometry, Staining

Journal: Scientific Reports

Article Title: The cytoprotective role of DJ-1 and p45 NFE2 against human primary alveolar type II cell injury and emphysema

doi: 10.1038/s41598-018-21790-3

Figure Lengend Snippet: p45 NFE2 interaction with DJ-1 in ATII cells and lung tissue obtained from non-smokers (NS), smokers (SM) and patients with emphysema (E). DJ-1 was co-immunoprecipitated in freshly isolated ATII cells ( A ) or lung tissue ( B ) followed by Western blotting analysis to determine its interaction with p45 NFE2 or p18 NFE2 as described in Materials and Methods. Relative expression is also shown. ( C ) p45 NFE2 (green) and DJ-1 (red) expression in ATII cells identified using SP-A staining (violet) in lung tissue by immunohistofluorescence. Cell nuclei were stained with DAPI (blue). The strongest co-localization of p45 NFE2 and DJ-1 is indicated by white arrows. *Statistically significant difference ( p < 0.05). Data are shown as the mean (±s.e.m.).

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): MMP9, CD147, ADAMTSL-4, p45 NFE2, p18 NFE2, DJ-1, phosphoserine, phosphothreonine and phosphotyrosine.

Techniques: Immunoprecipitation, Isolation, Western Blot, Expressing, Staining, Immunohistofluorescence

Journal: Scientific Reports

Article Title: The cytoprotective role of DJ-1 and p45 NFE2 against human primary alveolar type II cell injury and emphysema

doi: 10.1038/s41598-018-21790-3

Figure Lengend Snippet: MMP9, CD147, cathepsin B, p18 NFE2, p45 NFE2 and ADAMTSL-4 expression in wild-type and DJ-1 KO mice. Wild-type (WT) ( A ) and DJ-1 KO mice ( B ) were exposed to 150 mg/m 3 cigarette smoke (CS) for 2 h/day for 3 weeks as described in Materials and Methods section. Protein levels were analyzed in lung tissue by Western blotting. Lane 1 – WT mice, Lane 2 – WT + CS, Lane 3 – DJ-1 KO mice, Lane 4 – DJ-1 KO mice + CS. Relative expression is also shown. ( C ) p45 NFE2 expression (green) in ATII cells identified using SP-A antibody (violet) in lung tissue obtained from wild-type and DJ-1 KO mice by immunohistofluorescence. Cell nuclei were stained with DAPI (blue). ( D , E ) MMP9, CD147, cathepsin B, p45 NFE2 and ADAMTSL-4 mRNA expression in lung tissue from wild-type ( D ) and DJ-1 KO ( E ) mice by RT-PCR. The strongest p45 NFE2 expression is indicated by white arrows. *Statistically significant difference in comparison with control ( p < 0.05). Data are shown as the mean (±s.e.m.).

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): MMP9, CD147, ADAMTSL-4, p45 NFE2, p18 NFE2, DJ-1, phosphoserine, phosphothreonine and phosphotyrosine.

Techniques: Expressing, Western Blot, Immunohistofluorescence, Staining, Reverse Transcription Polymerase Chain Reaction

Journal: Scientific Reports

Article Title: The cytoprotective role of DJ-1 and p45 NFE2 against human primary alveolar type II cell injury and emphysema

doi: 10.1038/s41598-018-21790-3

Figure Lengend Snippet: The role of p45 NFE in ATII cells in non-smokers, smokers and emphysema patients.

Article Snippet: The following antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA): MMP9, CD147, ADAMTSL-4, p45 NFE2, p18 NFE2, DJ-1, phosphoserine, phosphothreonine and phosphotyrosine.

Techniques:

Journal: Journal of Oncology

Article Title: miR-23b-3p Inhibits the Oncogenicity of Colon Adenocarcinoma by Directly Targeting NFE2L3

doi: 10.1155/2021/8493225

Figure Lengend Snippet: NFE2L3 is a direct target gene of miR-23b-3p. (a) The miRanda database showing the binding sites of miR-23b-3p with 3ʹ-UTR of NFE2L3 and the mutant sequence. (b) Luciferase reporter assay was used to determine whether the binding between miR-23b-3p and 3ʹ-UTR of NFE2L3 affected the expression of NFE2L 3. (c) The mRNA level of NFE2L3 was measured using qRT-PCR analysis after transfecting COAD cells with miR-23b-3p mimics or NC. ∗ P < 0.05 vs. NC group. (d) Spearman's correlation analysis correlation between miR-23b-3p and NFE2L3 mRNA levels in COAD tissues of Figure 1(a) ( n = 23). (e, f) Expression profiles of NFE2L3 in COAD at different stages and normal colon tissues using the databases in the UALCAN platform. The results are expressed as the mean ± SD from three independent experiments. ∗ P < 0.05 vs. normal colon tissues.

Article Snippet: Then, the membranes were blocked with 5% (w/v) nonfat milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) at room temperature and incubated with a primary antibody against NFE2L3 (Proteintech, Rosemont, IL, USA) at 4°C overnight (diluted at 1 : 800).

Techniques: Binding Assay, Mutagenesis, Sequencing, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR

Journal: Journal of Oncology

Article Title: miR-23b-3p Inhibits the Oncogenicity of Colon Adenocarcinoma by Directly Targeting NFE2L3

doi: 10.1155/2021/8493225

Figure Lengend Snippet: Restoration of NFE2L3 abrogates the antitumor effects of miR-23b-3p in COAD cell. MiR-23 b-3p mimics were cotransfected into LoVo cells with pcDNA 3.1-NFE2L3 or pcDNA 3.1. (a–c) The proliferation was determined using the CCK-8, cell colony formation, and EdU assays. (d, e) The migration and invasion were determined using the Transwell assay. (f) Cell apoptosis was determined using flow cytometry. The results are expressed as the mean ± SD from three independent experiments. ∗ P < 0.05 miR-23b-3p mimics + Ov-NFE2L3 vs. miR-23b-3p mimics + Ov-Ctrl.

Article Snippet: Then, the membranes were blocked with 5% (w/v) nonfat milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) at room temperature and incubated with a primary antibody against NFE2L3 (Proteintech, Rosemont, IL, USA) at 4°C overnight (diluted at 1 : 800).

Techniques: CCK-8 Assay, Migration, Transwell Assay, Flow Cytometry

Journal: Journal of Oncology

Article Title: miR-23b-3p Inhibits the Oncogenicity of Colon Adenocarcinoma by Directly Targeting NFE2L3

doi: 10.1155/2021/8493225

Figure Lengend Snippet: Expression of miR-23b-3p inhibits COAD growth in vivo. (a) Tumor volume was measured every seven days. (b) Tumors were isolated after 28 days of the injection of LoVo cells, and images of representative tumors were captured. The tumor volume and weight were calculated ( n = 6). (c) The mRNA of miR-23b-3p in isolated tumors were measured by qRT-PCR analysis. (d) The expression of NFE2L3 was detected by Western blot analysis. ∗ P < 0.05 vs. NC.

Article Snippet: Then, the membranes were blocked with 5% (w/v) nonfat milk in Tris-buffered saline containing 0.1% Tween 20 (TBST) at room temperature and incubated with a primary antibody against NFE2L3 (Proteintech, Rosemont, IL, USA) at 4°C overnight (diluted at 1 : 800).

Techniques: Expressing, In Vivo, Isolation, Injection, Quantitative RT-PCR, Western Blot

Journal: Neural Regeneration Research

Article Title: Electroacupuncture alleviates cerebral ischemia and reperfusion injury via modulation of the ERK1/2 signaling pathway

doi: 10.4103/1673-5374.187041

Figure Lengend Snippet: Effects of EA on mRNA expression levels of Bax, Bcl-2, GCS and Nrf2 in the ischemic cortex after MCAO. Bax (A), Bcl-2 (B), GCSh (D), GCSl (E) and Nrf2 (F) mRNA expression levels, as assessed by real-time PCR. The Bax/Bcl-2 ratio is shown in C. Data ( n = 7 per group in A–C, n = 5 per group in D–F) are expressed as the mean ± SEM. Statistical significance was determined using analysis of variance followed by Tukey’s multiple comparison test for multiple comparisons. * P < 0.05, ** P < 0.01, vs . sham (sham-operation) group; # P < 0.05, vs . MCAO group; † P < 0.05, vs. EA (MCAO + EA) group. MCAO: Middle cerebral artery occlusion; EA: electroacupuncture; EA plus PD98059: MCAO + EA + PD98059; GCSh: gamma-glutamylcysteine synthetase heavy subunit; GCSl: gamma-glutamylcysteine synthetase light subunit; Nrf2: nuclear factor erythroid 2-related factor 2.

Article Snippet: Then, the sections were incubated with rabbit anti-rat GCS monoclonal antibody (1:100; BA1627, Wuhan Boster Biotechnology Company, Wuhan, Hubei Province, China) or rabbit anti-rat Nrf2 monoclonal antibody(1:150; PB0327, Wuhan Boster Biotechnology Company), as the primary antibody, and then with horseradish peroxidase-conjugated goat anti-rabbit IgG, as the secondary antibody (1:100; BA1055, Wuhan Boster Biotechnology Company).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Comparison

Journal: Neural Regeneration Research

Article Title: Electroacupuncture alleviates cerebral ischemia and reperfusion injury via modulation of the ERK1/2 signaling pathway

doi: 10.4103/1673-5374.187041

Figure Lengend Snippet: Effects of EA on GCS and Nrf2 immunoreactivities in the ischemic cortex. (A) Immunohistochemical staining for GCS and Nrf2 in the cortex in the different groups (× 400) ( n = 5 per group). (B, C) Mean optical densities of GCS and Nrf2-immunoreactive cells. Data are expressed as the mean ± SEM. Paired t -test was used to compare two groups, while analysis of variance followed by Tukey’s multiple comparisons test was used for multiple comparisons. ** P < 0.01, vs . sham (sham-operation) group; ## P < 0.01, vs . MCAO group; †† P < 0.01, vs . EA (MCAO + EA) group. EA: Electroacupuncture; GCS: glutamylcysteine synthetase; Nrf2: nuclear factor erythroid 2-related factor 2; MCAO: middle cerebral artery occlusion; EA plus PD98059: MCAO + EA + PD98059.

Article Snippet: Then, the sections were incubated with rabbit anti-rat GCS monoclonal antibody (1:100; BA1627, Wuhan Boster Biotechnology Company, Wuhan, Hubei Province, China) or rabbit anti-rat Nrf2 monoclonal antibody(1:150; PB0327, Wuhan Boster Biotechnology Company), as the primary antibody, and then with horseradish peroxidase-conjugated goat anti-rabbit IgG, as the secondary antibody (1:100; BA1055, Wuhan Boster Biotechnology Company).

Techniques: Immunohistochemical staining, Staining