Journal: Biomedicines

Article Title: Consequences of the Lack of TNFR1 in Ouabain Response in the Hippocampus of C57BL/6J Mice

doi: 10.3390/biomedicines10112937

Figure Lengend Snippet: Effect of ouabain and the lack of TNFR1 in TNF levels, TNFR2 expression, and TACE/ADAM17 activity. ( A ) TNF levels in serum were not changed by ouabain treatment or the TNFR1 gene. ( B ) TNFR1 KO mice presented the same TNF levels as WT mice in hippocampus. ( C ) Densitometric analysis and representative Western blotting of TNFR2 membrane expression. There was an increase in TNFR2 expression in the membrane enriched fraction for the gene factor [F (1, 30) = 5.591]. ( D ) The activity of TACE/ADAM 17 was raised for ouabain treatment [F (1, 16) = 4563]. Results are expressed in pg/mL for serum samples ( n = 11, N = 3) and pg/υg (mean ± SEM) in hippocampal samples ( n = 4, N = 2) for TNF measurements, in control ratio for TNFR2 expression ( n = 5, N = 2), and ng/mg in TACE/ADAM 17 activity ( n = 5, N = 1). Two-way ANOVA was performed for the comparison followed by Tukey´s post-test.

Article Snippet: A mouse TACE/ADAM17 kit was used to measure ADAM17 activity (BT Lab).

Techniques: Expressing, Activity Assay, Western Blot, Membrane, Control, Comparison

Journal: The Journal of Neuroscience

Article Title: l -3- n -Butylphthalide Improves Cognitive Impairment and Reduces Amyloid-β in a Transgenic Model of Alzheimer's Disease

doi: 10.1523/JNEUROSCI.0340-10.2010

Figure Lengend Snippet: Primary antibodies used in this study

Article Snippet: TACE , C terminal of human TACE , Rabbit , 1:200 , WB , ProSci.

Techniques: IF-P

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: Expression of miR-145, ADAM17, ACE2, and Mas receptor in the aortic arteries of hypertensive rats. 22 male Wistar rats were selected randomly to receive standard diet or high-sucrose/high-fat diet for 30 weeks, the thoracic aorta was collected, the expression of ACE2 was detected by Western blotting, and the expression of MASR, miR-145, and ADAM17 was detected by qPCR. (a) Quantification of ACE2 expression from panel (b) data was expressed after normalization to the β -actin. (b) Representative figures from Western blotting of ACE2. (c–e) Quantification of mRNA expression of MASR, miR-145, and ADAM17 (qPCR). ∗∗ P < 0.01 between groups and ∗ P < 0.05 between groups.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: Expressing, Western Blot

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: miR-145 augments Ang II-induced ACE2-Ang-(1–7)-Mas axis activation and ADAM17 expression in VSMCs. VSMCs were treated with control, Ang II (1 μ M), miR-145 mimic (100 nM), or miR-145 (100 nM) inhibitor alone or in combination for 48 hours. (a) Concentration of Ang-(1–7) in the supernatant (ELISA). (b, c) ACE2 and MASR expression from panel (e) and data are expressed as the fold of the GAPDH. (d) ADAM17 expression in groups from panel (f). (e, f) Representative figures of ACE2, MASR, and ADAM17 expression in VSMCs (Western blotting). ∗∗ P < 0.01 vs. control group; ∗ P < 0.05 vs. control group; ## P < 0.01 vs. Ang II group; and # P < 0.05 vs. Ang II group. All the data are expressed as mean ± SEM from three independent experiments. miRNA NC, negative control miRNA.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: Activation Assay, Expressing, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Negative Control

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: ADAM17 siRNA reversed phenotype transition induced by miR-145 in vitro. VSMCs were treated with miR-145 inhibitor in the presence or absence of ADAM17 siRNA (100 nM) as indicated for 48 hours. The expression of α -SMA (a), SM22 α (b), OPN (c), EREG (d), and MMP2 (e) protein was detected by Western blotting. All the data were normalized to that of GAPDH. (f) Representative figures of OPN, α -SMA, SM22 α , EREG, and MMP2 (Western blotting). (g) The luciferase reporter assay is shown. Cells were transfected with a reporter vector psiCHECK-2-ADAM17 3′-UTR plus either miR-145-5p or the negative control. ∗∗ P < 0.01 vs. control group or miR-145-5p NC and ## P < 0.01 vs. miR-145 inhibitor group. All the data are expressed as mean ± SEM of three independent experiments. NC siRNA, negative control siRNA. WT, wide type. MT, mutant type.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: In Vitro, Expressing, Western Blot, Luciferase, Reporter Assay, Transfection, Plasmid Preparation, Negative Control, Control, Mutagenesis

Journal: International Journal of Hypertension

Article Title: miR-145 Alleviates Smooth Muscle Cell Phenotype Transition via ADAM17-Mediated ACE2 Shedding

doi: 10.1155/2023/9497716

Figure Lengend Snippet: ADAM17 mediated miR-145-induced effect by regulating ACE2-Ang-(1–7)-Mas axis in vitro. VSMCs were treated with miR-145 inhibitor in the presence or absence of ADAM17 siRNA (100 nM) as indicated for 48 hours. (a) Concentration of Ang-(1–7) in the supernatant (ELISA). (b) Representative figures of ACE2 and MASR (Western blotting). Quantification of ACE2 (c) and MASR (d) expression determined by Western blotting, and the data were normalized to that of GAPDH. ∗∗ P < 0.01 vs. control group; ∗ P < 0.05 vs. control group; and ## P < 0.01 vs. miR-145 inhibitor group. All the data are expressed as mean ± SEM of three independent experiments. NC siRNA, negative control siRNA.

Article Snippet: Antibodies against angiotensin converting enzyme 2 (ACE2) (21115-1-AP), Mas receptor (20080-1-AP), osteopontin (OPN) (22952-1-AP), α -SMA (Proteintech, USA, 1 : 2000), SM22a (10493-1-AP), ADAM17 (20259-1-AP), EREG (CSB-PA189260), MMP2 (10373-2-AP) and GAPDH (10494-1-AP), and HRP goat anti-rabbit IgG (SA00001-2) were purchased from Proteintech (Proteintech, USA).

Techniques: In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Control, Negative Control

Journal: Brain

Article Title: Cerebrovascular lesions induce transient ?-amyloid deposition

doi: 10.1093/brain/awr300

Figure Lengend Snippet: Transient middle cerebral artery occlusion did not affect proteins related to generation or degradation of β-amyloid. Representative examples of the limited effect of middle cerebral artery occlusion on proteins related to β-amyloid production or degradation using immunohistochemistry in post-mortem tissue sections. BACE, presenilin 1 and APP immunoreactivity was observed in the surrounding of senile plaques (not labelled), whereas neprilysin and insulin degrading enzyme signal was detected in neurons. No differences were observed in BACE (A and B), presenilin 1 (C and D), APP (E and F), neprilysin (G and H) or insulin degrading enzyme (I and J) (top: ipsilateral hemisphere; bottom: contralateral hemisphere). Scale bar = 200 μm.

Article Snippet: Reagents Texas Red® dextran of 70 000 D molecular weight was obtained from Molecular Probes; methoxy-XO4 was a gift from Dr Klunk (University of Pittsburgh); Ketamine HCl and xylazine were obtained from Phoenix Pharmaceuticals, anti-β-amyloid antibody (10D5) (developed in mouse) was a gift from Elan Pharmaceuticals; anti-β-amyloid-converting enzyme (BACE) antibody (developed in rabbit) was obtained from ProSci Incorporated; anti-amyloid precursor protein (APP) antibody (developed in rabbit) was obtained from Calbiochem; anti-insulin degrading enzyme (developed in goat) was obtained from Santa Cruz; and anti-neprilysin (developed in rat) was obtained from R&D Systems.

Techniques: Immunohistochemistry

Journal: Frontiers in bioscience (Landmark edition)

Article Title: MiR-145 Alleviates Sepsis-Induced Inflammatory Responses and Organ Injury by Targeting ADAM17.

doi: 10.31083/j.fbl2901044

Figure Lengend Snippet: Fig. 1. Lipopolysaccharide (LPS) reduces miR-145 expression, overexpression of miR-145 downregulated ADAM17 expression

Article Snippet: The membranes were incubated with antibodies against ADAM17 (sc-390859, mouse monoclonal antibody, 1:800, Santa Cruz Biotechnology, Santa Cruz, CA, USA; A00604, rabbit polyclonal antibody, 1:800, Boster Biological Technology, China) or β-actin (sc-47778, mouse monoclonal antibody, 1:4000, Santa Cruz Biotechnology, USA) at 4 °C overnight.

Techniques: Expressing, Over Expression

Journal: Frontiers in bioscience (Landmark edition)

Article Title: MiR-145 Alleviates Sepsis-Induced Inflammatory Responses and Organ Injury by Targeting ADAM17.

doi: 10.31083/j.fbl2901044

Figure Lengend Snippet: Fig. 2. miR-145 alleviated LPS-induced endothelial inflammation by targeting ADAM17 in HUVECs. HUVECs were infected by LV-NC, LV-miR-145, LV-ADAM17-siRNA or the combined of LV-miR-145 and LV-ADAM17-siRNA. (A) Green fluorescence protein

Article Snippet: The membranes were incubated with antibodies against ADAM17 (sc-390859, mouse monoclonal antibody, 1:800, Santa Cruz Biotechnology, Santa Cruz, CA, USA; A00604, rabbit polyclonal antibody, 1:800, Boster Biological Technology, China) or β-actin (sc-47778, mouse monoclonal antibody, 1:4000, Santa Cruz Biotechnology, USA) at 4 °C overnight.

Techniques: Infection, Fluorescence

Journal: Frontiers in bioscience (Landmark edition)

Article Title: MiR-145 Alleviates Sepsis-Induced Inflammatory Responses and Organ Injury by Targeting ADAM17.

doi: 10.31083/j.fbl2901044

Figure Lengend Snippet: Fig. 3. Overexpression of miR-145 reduces expression of ADAM17, attenuates sepsis-induced inflammatory responses and acute lung injury. (A) Schematic of experimental design and time line. Polymicrobial sepsis model of mice was induced in C57BL/6 mice

Article Snippet: The membranes were incubated with antibodies against ADAM17 (sc-390859, mouse monoclonal antibody, 1:800, Santa Cruz Biotechnology, Santa Cruz, CA, USA; A00604, rabbit polyclonal antibody, 1:800, Boster Biological Technology, China) or β-actin (sc-47778, mouse monoclonal antibody, 1:4000, Santa Cruz Biotechnology, USA) at 4 °C overnight.

Techniques: Over Expression, Expressing

Journal: Frontiers in bioscience (Landmark edition)

Article Title: MiR-145 Alleviates Sepsis-Induced Inflammatory Responses and Organ Injury by Targeting ADAM17.

doi: 10.31083/j.fbl2901044

Figure Lengend Snippet: Fig. 4. MiR-145 agomir reduces expression of ADAM17, attenuates sepsis-induced acute kidney injury and offers survival benefit.

Article Snippet: The membranes were incubated with antibodies against ADAM17 (sc-390859, mouse monoclonal antibody, 1:800, Santa Cruz Biotechnology, Santa Cruz, CA, USA; A00604, rabbit polyclonal antibody, 1:800, Boster Biological Technology, China) or β-actin (sc-47778, mouse monoclonal antibody, 1:4000, Santa Cruz Biotechnology, USA) at 4 °C overnight.

Techniques: Expressing