Journal: Cancers

Article Title: Novel Insights into the Antagonistic Effects of Losartan against Angiotensin II/AGTR1 Signaling in Glioblastoma Cells

doi: 10.3390/cancers13184555

Figure Lengend Snippet: Aromatase expression in SVG p12 normal glial cells and U-87 MG and T98G glioblastoma cells. ( A ) Real-time RT-PCR for CYP19A1 in SVG p12 normal glial cells, U-87 MG, and T98G glioblastoma cells; mRNA is shown relative to SVG p12 normal glial cells. ( B ) Immunoblotting showing Arom protein expression. β-actin was used as a control for equal loading and transfer. The histograms represent the mean average ± SD of three separate experiments in which band intensities were evaluated in terms of optical density arbitrary unit and expressed as fold change over SVG p12 for U-87 MG and T98G. ( C ) Immunofluorescence of Arom in SVG p12 normal glial cells, U-87 MG, and T98G glioblastoma cells. DAPI staining for nuclear detection. Scale bars = 5 µm. Original magnification, ×100. Data are expressed as means ± SD of three different experiments, each performed in triplicate. *** p < 0.001.

Article Snippet: The plasmids pEZX-PL01 containing human aromatase promoter region (pI.f −875/−1) and the 5′-deleted promoter fragment pI.f −674/−1 plasmids were generated by GeneCopeia (Rockville, MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Staining

Journal: Cancers

Article Title: Novel Insights into the Antagonistic Effects of Losartan against Angiotensin II/AGTR1 Signaling in Glioblastoma Cells

doi: 10.3390/cancers13184555

Figure Lengend Snippet: Aromatase and clinical outcomes in GBM patients. Kaplan–Meier survival analysis relating CYP19A1 levels and overall survival (OS) in GBM patients (TCGA dataset).

Article Snippet: The plasmids pEZX-PL01 containing human aromatase promoter region (pI.f −875/−1) and the 5′-deleted promoter fragment pI.f −674/−1 plasmids were generated by GeneCopeia (Rockville, MD, USA).

Techniques:

Journal: Cancers

Article Title: Novel Insights into the Antagonistic Effects of Losartan against Angiotensin II/AGTR1 Signaling in Glioblastoma Cells

doi: 10.3390/cancers13184555

Figure Lengend Snippet: Effects of Ang II and LOS on aromatase expression and activity in U-87 MG and T98G glioblastoma cells. ( A , B ) Real-time RT-PCR and immunoblotting assay for CYP19A1 mRNA and protein expression in U-87 MG and T98G glioblastoma cells, treated with vehicle (−) or the Ang II 0.1, 5, and 10 µM for 24 h. GAPDH was used as a loading control. The histograms represent the mean average ± SD of three separate experiments in which band intensities were evaluated in terms of optical density arbitrary unit and expressed as fold change over vehicle (−) for Ang II treatment. ( C , D ) CYP19A1 mRNA and protein expression in U-87 MG and T98G glioblastoma cells treated with vehicle (−) and Ang II 5 µM alone or in combination with LOS 5 µM for 24 h. The histograms represent the mean average ± SD of three separate experiments in which band intensities were evaluated in terms of optical density arbitrary unit and expressed as fold change over vehicle (−) for Ang II treatment or fold change over Ang II for Ang II in combination with LOS, 5 µM. GAPDH was used as a loading control. ( E , G ) Aromatase activity in U-87 MG and T98G glioblastoma cells treated with vehicle (−) and Ang II 5 µM alone or in combination with LOS 5 µM for 24 h. ( F , H ) ELISA for Estradiol secretion in U-87 MG and T98G glioblastoma cells treated with vehicle (−) and Ang II 5 µM alone or in combination with LOS 5 µM for 24 h. Data represent the mean ± SD of three different experiments, each performed in triplicate. ** p < 0.01; *** p < 0.001.

Article Snippet: The plasmids pEZX-PL01 containing human aromatase promoter region (pI.f −875/−1) and the 5′-deleted promoter fragment pI.f −674/−1 plasmids were generated by GeneCopeia (Rockville, MD, USA).

Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Novel Insights into the Antagonistic Effects of Losartan against Angiotensin II/AGTR1 Signaling in Glioblastoma Cells

doi: 10.3390/cancers13184555

Figure Lengend Snippet: Aromatase promoters pI.f/I.4 in U-87 MG glioblastoma cells are stimulated upon Ang II exposure. ( A , B ) Upper panel, schematic map of the pI.f and pI.4 aromatase constructs. U-87 MG glioblastoma cells were transiently transfected with the reported constructs and treated with vehicle (−) or Ang II 5 µM, alone or in combination with LOS 5 µM for 24 h. ( C , D ) U-87 MG glioblastoma cells were treated in the presence of vehicle (−) or Ang II 5 µM for 3 h. Nuclear extracts were incubated with a biotinylated oligonucleotide containing the AP-1 or AP-1 mutant site in the aromatase promoters pI.f (left panel) or with a biotinylated oligonucleotide containing the STAT3 or STAT3 mutant site in the aromatase promoters pI.4 (right panel) and subjected to DNA affinity precipitation assay. Specifically bound proteins were subjected to Western blotting analysis. The specificity of the binding was tested by loading the unbound fraction (Negative Control). U-87 MG glioblastoma cells nuclear extracts were used as positive control. The histograms represent the mean average ± SD of three separate experiments in which band intensities were evaluated in terms of optical density arbitrary unit and expressed as fold change over vehicle (−) for Ang II treatment. ( E , F ) U-87 MG glioblastoma cells were treated in the presence of vehicle (−) or Ang II 5 µM for 3 h, then cross-linked with formaldehyde and lysed. The precleared chromatin was immunoprecipitated with anti-c-Jun or anti-RNA Pol II (left panel) or with anti-STAT3 and anti-RNA Pol II (the right panel). The 5′flanking sequence of the CYP19A1 gene was detected by real-time PCR with specific primers to amplify aromatase promoter sequence, including the AP-1 and STAT3 sites. Input DNA was amplified as loading controls. Data are expressed as means ± SD of three different experiments, each performed in triplicate. * p < 0.05; *** p < 0.001.

Article Snippet: The plasmids pEZX-PL01 containing human aromatase promoter region (pI.f −875/−1) and the 5′-deleted promoter fragment pI.f −674/−1 plasmids were generated by GeneCopeia (Rockville, MD, USA).

Techniques: Construct, Transfection, Incubation, Mutagenesis, Affinity Precipitation, Western Blot, Binding Assay, Negative Control, Positive Control, Immunoprecipitation, Sequencing, Real-time Polymerase Chain Reaction, Amplification

Journal: bioRxiv

Article Title: Transgenerational inheritance of betaine-induced epigenetic alterations in estrogen-responsive IGF-2/IGFBP2 genes in rat hippocampus

doi: 10.1101/640524

Figure Lengend Snippet: Effect of maternal betaine supplementation during pregnancy and lactation on aromatase (CYP19A1) and estrogen receptor related gene (ERα, ERβ, ERRα, ERRβ and ERRγ) mRNA expressions, and the promoter methylation levels in F0, F1 and F2 generation rat offspring hippocampus. (A) CYP19A1 and estrogen receptor related gene mRNA expressions in F0 generation; (B) CYP19A1 and estrogen receptor related gene promoter methylation levels in F0 generation; (C) CYP19A1 protein expression in F0 generation; (D) CYP19A1 and estrogen receptor related gene mRNA expressions in F1 generation; (E) CYP19A1 and estrogen receptor related gene promoter methylation levels F1 generation; (F) CYP19A1 protein expression in F1 generation; (G) CYP19A1 and estrogen receptor related gene mRNA expressions in F2 generation; (H) CYP19A1 and estrogen receptor related gene promoter methylation levels F2 generation; (I) CYP19A1 protein expression in F2 generation. Values are means ± SEM, * P < 0.05, ** P <0.01, compared with control (n = 6).

Article Snippet: Western blot analysis for IGF2 (BS6632, Bioworld, diluted 1:1000), CYP19A1 (BS6580, Bioworld, diluted 1:1000), betaine homocysteine methyltransferase (BHMT) (15965-1-AP, Proteintech, diluted 1:500), was carried out according to the recommended protocols provided by the manufacturers, and β-actin (AC026, ABclonal, diluted 1:50,000), Tubulinα (BS1699, Bioworld, diluted 1:10,000) was used as loading control.

Techniques: Methylation, Expressing

Journal: Food Science and Technology Research

Article Title: Anti-adipogenic Effects of Polyphenol Extracts of Areca Flower Tea on 3T3-L1 Preadipocytes

doi: 10.3136/fstr.23.705

Figure Lengend Snippet: Fig. 5. Effects of different AFPE concentrations on the expression of PPARγ and C/EBPα in adipocytes. Preadipocyte, blank control; Adipocyte, control; Resveratrol, positive control. All data represent as means ± SD of three independent experiments. Mean values with different letters were determined to be significantly different by one-way ANOVA followed by Duncan’s test (P < 0.05).

Article Snippet: The mouse PPARγ ELISA kit and mouse C/EBPα ELISA kit were purchased from Cusabio Co. (Wuhan, China).

Techniques: Expressing, Control, Positive Control

Journal: Current Atherosclerosis Reports

Article Title: RNA Therapeutics: the Next Generation of Drugs for Cardiovascular Diseases

doi: 10.1007/s11883-022-01007-9

Figure Lengend Snippet: RNA therapeutics in clinical trials for cardiovascular disease

Article Snippet: ARO APOC3 , siRNA , Phase III; Phase II , Dyslipidemias; hypertriglyceridemia (II); hyperlipoproteinemia type I (III) , Arrowhead Pharmaceuticals , Reduce production of apolipoprotein C-III (apoC-III) to lower very-low-density lipoprotein , NCT04720534 NCT04998201 NCT05089084 , ARO-APOC3.

Techniques: Clinical Proteomics, Inhibition, Coagulation, Expressing, Control, Binding Assay, Over Expression