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ChIP assay

Yingfeng Yang, Fangyu Xie, Dandan Qin, Chen Zong, Feng Han, Zeqing Pu, Dong Liu, Xia Li, Yuchao Zhang, Yuantao Liu, Xiangdong Wang
AIMS: Our previous study showed that NR4A1 protects against oxidative stress-induced cell apoptosis. However, the targets downstream of NR4A1 are incompletely known. Glutathione peroxidase 1 (GPX1) is the most common antioxidant enzyme in the glutathione peroxidase class. In this study, we aimed to investigate whether GPX1 is a mediator of the protective effects of NR4A1 in pancreatic β cells. MAIN METHODS: A pancreatic β cell line, MIN6, was used to generate NR4A1 over-expression cell line...
April 17, 2018: Life Sciences
Alessandra Micera, Antonio Di Zazzo, Graziana Esposito, Rosa Longo, William Foulsham, Roberto Sacco, Roberto Sgrulletta, Stefano Bonini
Purpose: We characterize age-associated alterations in the expression of inflammatory mediators and tissue remodeling factors in human tears. Methods: A total of 75 consecutive volunteers (32 male/44 female; 19-93 years) underwent clinical assessment of ocular surface status, ocular surface disease index (OSDI) grading and tear sampling. The volunteers were categorized into three groups: young (18-40 years), middle-aged (41-60 years), and old (>60 years). Total protein profiles and chip-based protein array evaluations were conducted to investigate the expression of 60 potential candidates, including pro-/anti-inflammatory mediators and tissue remodeling factors...
April 1, 2018: Investigative Ophthalmology & Visual Science
Adrian C Robu, Laurentiu Popescu, Daniela G Seidler, Alina D Zamfir
Fibroblast growth factor-2 (FGF-2) is involved in wound healing and embryonic development. Glycosaminoglycans (GAGs), the major components of the extracellular matrix (ECM), play fundamental roles at this level. FGF-GAG noncovalent interactions are in the focus of research, due to their influence upon cell proliferation and tissue regeneration. Lately, high resolution mass spectrometry (MS) coupled with chip-nanoelectrospray (nanoESI) contributed a significant progress in glycosaminoglycomics by discoveries related to novel species and their characterization...
April 20, 2018: Journal of Mass Spectrometry: JMS
Miranda L Xu, Wilson K W Luk, Cathy W C Bi, Etta Y L Liu, Kevin Q Y Wu, Ping Yao, Tina T X Dong, Karl W K Tsim
Acetylcholinesterase (AChE; EC is known to hydrolyze acetylcholine at cholinergic synapses. In mammalian erythrocyte, AChE exists as a dimer (G2 ) and is proposed to play role in erythropoiesis. To reveal the regulation of AChE during differentiation of erythroblast, erythroblast-like cells (TF-1) were induced to differentiate by application of erythropoietin (EPO). The expression of AChE was increased in parallel to the stages of differentiation. Application of EPO in cultured TF-1 cells induced transcriptional activity of ACHE gene, as well as its protein product...
April 20, 2018: Journal of Neurochemistry
Amber M Messmer, Jong S Leong, Eric B Rondeau, Anita Mueller, Cody A Despins, David R Minkley, Matthew P Kent, Sigbjørn Lien, Brad Boyce, Diane Morrison, Mark D Fast, Joseph D Norman, Roy G Danzmann, Ben F Koop
Antiparasitic drugs such as emamectin benzoate (EMB) are relied upon to reduce the parasite load, particularly of the sea louse Lepeophtheirus salmonis, on farmed salmon. The decline in EMB treatment efficacy for this purpose is an important issue for salmon producers around the world, and particularly for those in the Atlantic Ocean where widespread EMB tolerance in sea lice is recognized as a significant problem. Salmon farms in the Northeast Pacific Ocean have not historically experienced the same issues with treatment efficacy, possibly due to the relatively large population of endemic salmonid hosts that serve to both redistribute surviving lice and dilute populations potentially under selection by introducing naïve lice to farms...
April 16, 2018: Marine Genomics
Desheng Kong, Linlin Zhao, Lili Sun, Shengjin Fan, Huibo Li, Yanqiu Zhao, Zhibo Guo, Leilei Lin, Lin Cui, Ke Wang, Wenjia Chen, Yihui Zhang, Jin Zhou, Yinghua Li
Dickkopf-3 (DKK3) is frequently down-regulated by promoter hypermethylation and is closely associated with a poor prognosis in many cancers. Our previous studies have shown that miR-708 down-regulates DKK3 at the post-transcriptional level in B-ALL. However, whether transcriptional mechanisms lead to DKK3 silencing remains unclear. Here, we analysed the promoter regions of DKK3 by bioinformatics and found binding sites for MYCN. A dual-luciferase reporter gene assay and ChIP experiments revealed that MYCN negatively regulates DKK3 at the transcriptional level in B-ALL cell lines, and using bisulphite sequencing PCR, we affirmed that MYCN has no effect on the methylation of the DKK3 promoter...
April 19, 2018: Journal of Cellular and Molecular Medicine
Chih-Ming Weng, Chun-Hua Wang, Meng-Jung Lee, Jung-Re He, Hsin-Yu Huang, Ming-Wei Chao, Kian Fan Chung, Han-Pin Kuo
BACKGROUND: Exposure to environmental pollutants promotes Th2 cell responses. Aryl hydrocarbon receptor (AhR) activation aggravates allergic responses. Epithelium-derived thymic stromal lymphopoietin (TSLP), interleukin (IL)-25 and IL-33 are implicated in the dysregulation of Th2 immune responses in severe allergic asthma. METHODS: Bronchial biopsies of 28 allergic severe asthma and 6 mild asthma subjects from highly polluted areas were analyzed for AhR nuclear translocation (NT), cytokine expression and gene activation...
April 19, 2018: Allergy
Maxime Huet, Myriam Cubizolles, Arnaud Buhot
Pre-transfusion bedside compatibility test is mandatory to check that the donor and the recipient present compatible groups before any transfusion is performed. Although blood typing devices are present on the market, they still suffer from various drawbacks, like results that are based on naked-eye observation or difficulties in blood handling and process automation. In this study, we addressed the development of a red blood cells (RBC) agglutination assay for point-of-care blood typing. An injection molded microfluidic chip that is designed to enhance capillary flow contained anti-A or anti-B dried reagents inside its microchannel...
April 19, 2018: High-throughput
Fangjun Yu, Tianpeng Zhang, Lianxia Guo, Baojian Wu
Organic anion transporter 2 (OAT2/SLC22A7) is an uptake transporter that plays an important role in drug disposition. Here, we investigate a potential role of liver receptor homolog-1 (Lrh-1) in regulation of Oat2 and docetaxel pharmacokinetics. Hepatoma cells (Hepa1-6 and HepG2 cells) were transfected with Lrh-1/LRH-1 expression vector or siRNA. The relative mRNA and protein levels of Oat2/OAT2 in the cells or livers of Lrh-1hep-/- mice were determined by qPCR and Western blotting, respectively. Transcriptional regulation of Oat2/OAT2 by Lrh-1/LRH-1 were investigated using luciferase reporter, mobility shift, and chromatin immunoprecipitation (ChIP) assays...
April 18, 2018: Drug Metabolism and Disposition: the Biological Fate of Chemicals
Silvia Armelloni, Masami Ikehata, Deborah Mattinzoli, Min Li, Carlo Maria Alfieri, Mariapia Rastaldi, Piergiorgio Messa
Background/Aims The research of genes implicated in kidney glomerular function, eliciting cell fate program, is always at the forefront in nephrological studies. Several neurological molecules have been recently the object of study not only for their involvement in the central nervous system differentiation but also for their importance in the functionality of other organs and for mature phenotype, as in kidney. NeuroD, in CNS, is related to two functional roles, the early survival and the differentiation. The aim of our study was to ascertain the presence of NeuroD transcription factor in glomeruli and to understand which targets and mechanisms NeuroD controls...
April 13, 2018: Cellular Physiology and Biochemistry
Jinpin Li, Di Qiu, Zezhi Chen, Weiwei Du, Jingli Liu, Xuean Mo
Myasthenia gravis (MG) is a B cell-mediated and T cell-dependent autoimmune disease. Thymic hyperplasia has great significance for MG pathogenesis and treatment. MicroRNAs (miRNAs) are a newly recognized type of gene expression regulatory factor that regulate gene expression at the post-transcriptional level. Additionally, miRNAs are involved in immune regulation of the thymus and the occurrence and development of autoimmune diseases. In this study, we found 33 miRNAs that were significantly dysregulated in thymic tissues from MG patients with thymus hyperplasia (MGH) compared with thymic tissues from normal controls using a miRNA microarray chip...
April 4, 2018: Journal of Neuroimmunology
Azhar Jabareen, Manal Salman, Aya Abu-Jaafar, Mahmoud Huleihel
HTLV-1 and HIV-1 are retroviruses involved in different human diseases. However, following infection, these viruses inter into a latent state. Tax and Tat are regarded as trans-activators of HTLV-1 and HIV-1 respectively. As it known, during the latent state the infected cells contain low Tax and Tat protein levels, so the activation of these viruses must be independent of these proteins. Here we focus on exploring the mechanism of activation of these viruses by 12-O-tetradecanoylphorbol-13-acetate (TPA), which is a potent activator of protein kinase C (PKC) and considered as a stress-inducing agent...
April 13, 2018: Biochemical and Biophysical Research Communications
Qianyi Tao, Huanhuan Niu, Zhongyuan Wang, Wenhui Zhang, Hu Wang, Shenhao Wang, Xian Zhang, Zheng Li
In plants, unisexual flowers derived from developmental sex determination form separate stamens and pistils that facilitate cross pollination. In cucumber and melon, ethylene plays a key role in sex determination. Six sex determination-related genes have been identified involving in ethylene biosynthesis in these Cucumis species. The interactions among these genes are thought to involve ethylene signaling; however, the detailed regulatory mechanism remains unknown. In this study, hormone treatment and qPCR assays were used to confirm expression of these sex determination-related genes in cucumber and melon is ethylene sensitive...
April 5, 2018: Journal of Experimental Botany
Zhijia Tan, Ben Niu, Kwok Yeung Tsang, Ian G Melhado, Shinsuke Ohba, Xinjun He, Yongheng Huang, Cheng Wang, Andrew P McMahon, Ralf Jauch, Danny Chan, Michael Q Zhang, Kathryn S E Cheah
The growth plate mediates bone growth where SOX9 and GLI factors control chondrocyte proliferation, differentiation and entry into hypertrophy. FOXA factors regulate hypertrophic chondrocyte maturation. How these factors integrate into a Gene Regulatory Network (GRN) controlling these differentiation transitions is incompletely understood. We adopted a genome-wide whole tissue approach to establish a Growth Plate Differential Gene Expression Library (GP-DGEL) for fractionated proliferating, pre-hypertrophic, early and late hypertrophic chondrocytes, as an overarching resource for discovery of pathways and disease candidates...
April 16, 2018: PLoS Genetics
Rob J A Verhoeven, Shuang Tong, Jingfeng Zong, Yixin Chen, Sai-Wah Tsao, Jianji Pan, Honglin Chen
Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is one of the few viral proteins expressed by EBV in nasopharyngeal carcinoma (NPC), most likely because of its essential role in maintaining the viral genome in EBV-infected cells. In NPC, EBNA1 expression is driven by the Bam HI-Q promoter (Qp), which is regulated by both cellular and viral factors. We previously determined that the expression of another group of EBV transcripts, Bam HI-A rightward transcripts (BARTs), is associated with constitutively activated nuclear factor-κB (NF-κB) signaling in NPC cells...
April 16, 2018: Cancers
Lanhuan Meng, Zhongqi Fan, Qiang Zhang, Cuicui Wang, Ying Gao, Yikang Deng, Benzhong Zhu, Hongliang Zhu, Chen Jianye, Wei Shan, Xunren Yin, Silin Zhong, Donald Grierson, Cai-Zhong Jiang, Yunbo Luo, Da-Qi Fu
Chloroplast development and chlorophyll(Chl)metabolism in unripe tomato contribute to the growth and quality of the fruit. However, the mechanism is poorly understood. In this work, we initially studied seven homeobox-containing transcription factors (TFs) with specific ripening-associated expression patterns by virus-induced gene silencing (VIGS) technology and found that inhibiting the expression of one of these TFs, BEL1-LIKE HOMEODOMAIN11 (SlBEL11), significantly increased Chl levels in unripe tomato fruit...
April 16, 2018: Plant Journal: for Cell and Molecular Biology
Anna M Chiarella, Austin L Quimby, Marjan Mehrab-Mohseni, Brian Velasco, Sandeep K Kasoji, Ian Davis, Paul A Dayton, Nathaniel A Hathaway, Samantha Pattenden
One of the most sensitive, time consuming, and variable steps of chromatin immunoprecipitation (ChIP) is chromatin sonication. Traditionally, this process can take hours to properly sonicate enough chromatin for multiple ChIP assays. Further, the length of sheared DNA is often inconsistent. In order to faithfully measure chemical and structural changes at the chromatin level, sonication needs to be reliable. Thus, chromatin fragmentation by sonication represents a significant bottleneck to downstream quantitative analysis...
April 16, 2018: Biochemistry
Ge Zi Li, Zhan Qi Wang, Kengo Yokosho, Bing Ding, Wei Fan, Qiao Qiao Gong, Gui Xin Li, Yun Rong Wu, Jian Li Yang, Jian Feng Ma, Shao Jian Zheng
Whilst WRKY transcription factors are known to be involved in diverse plant responses to biotic stresses, their involvement in abiotic stress tolerance is poorly understood. OsFRDL4, encoding a citrate transporter, has been reported to be regulated by ALUMINUM (Al) RESISTANCE TRANSCRIPTION FACTOR 1 (ART1) in rice, but whether it is also regulated by other transcription factors is unknown. We define the role of OsWRKY22 in response to Al stress in rice by using mutation and transgenic complementation assays, and characterize the regulation of OsFRDL4 by OsWRKY22 via yeas one-hybrid, electrophoretic mobility shift assay and ChIP-quantitative PCR...
April 15, 2018: New Phytologist
Shweta Bhagwat, Shraddha Sontakke, Deekshith K, Priyanka Parte, Sameer Jadhav
Chemotaxis, as a mechanism for sperm guidance in vivo , is an enigma which has been difficult to demonstrate. To address this issue, various devices have been designed to study sperm chemotaxis in vitro . Limitations of traditional chemotaxis devices were related to the inability to maintain a stable concentration gradient as well as track single sperm over long times. Microfluidics technology, which provides superior control over fluid flow, has been recently used to generate stable concentration gradients for investigating the chemotactic behavior of several cell types including spermatozoa...
March 2018: Biomicrofluidics
Yohei Nakayama, Sari Matsui, Keisuke Noda, Mizuho Yamazaki, Yasunobu Iwai, Bernhard Ganss, Yorimasa Ogata
Amelotin (AMTN) is induced upon initiation of apoptosis by transforming growth factor beta1 (TGFβ1) and is mediated by Smad3 in gingival epithelial cells (GE1 cells). This upregulation of AMTN gene expression is temporary, and the mechanism responsible is still unclear. The present study investigated the transcriptional downregulation of TGFβ1-induced AMTN gene expression in GE1 cells during the progression of apoptosis. To examine time-dependent changes in the levels of AMTN, Smad3 and Bax mRNA induced by TGFβ1, real-time PCR analyses were performed...
April 16, 2018: Journal of Oral Science
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