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Melina Schellhorn et al.
Oncotarget, 6(36), 39342-39356 (2015-10-30)
The antitumorigenic mechanism of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib is still a matter of debate. Using lung cancer cell lines (A549, H460) and metastatic cells derived from a lung cancer patient, the present study investigates the impact of celecoxib
Laura M Wingler et al.
Cell, 176(3), 479-490 (2019-01-15)
The angiotensin II (AngII) type 1 receptor (AT1R) is a critical regulator of cardiovascular and renal function and is an important model for studies of G-protein-coupled receptor (GPCR) signaling. By stabilizing the receptor with a single-domain antibody fragment ("nanobody") discovered
Ester Falconer et al.
Nature protocols, 5(7), 1362-1377 (2010-07-03)
Previously, assays for sister chromatid segregation patterns relied on incorporation of 5-bromo-2'-deoxyuridine (BrdU) and indirect methods to infer segregation patterns after two cell divisions. In this study, we describe a method to differentially label sister chromatids of mouse cells and
Siva S V P Sakamuri et al.
GeroScience, 40(3), 347-356 (2018-06-04)
Mitochondria play a critical role in the cardiomyocyte physiology by generating majority of the ATP required for the contraction/relaxation through oxidative phosphorylation (OXPHOS). Aging is a major risk factor for cardiovascular diseases (CVD) and mitochondrial dysfunction has been proposed as
Sang Hun Lee et al.
Neuron, 108(4), 676-690 (2020-09-07)
Amyloid precursor protein (APP) is associated with both familial and sporadic forms of Alzheimer's disease. Despite its importance, the role of APP family in neuronal function and survival remains unclear because of perinatal lethality exhibited by knockout mice lacking all
Liang Yang et al.
Archives of pharmacal research, 37(12), 1554-1559 (2014-08-15)
The present study aimed to develop an effective oral formulation of tranilast (TL), a poorly soluble anti-inflammatory drug, via the formation of drug complex with 3-aminopropyl functionalized magnesium phyllosilicate (AMP clay) and improve the pH-dependent drug dissolution and bioavailability of
Yusuke Tomina et al.
Bio-protocol, 8(5) (2018-03-30)
In this protocol, we introduce an effective method for voltage-sensitive dye (VSD) loading and imaging of leech ganglia as used in Tomina and Wagenaar (2017). Dissection and dye loading procedures are the most critical steps toward successful whole-ganglion VSD imaging.
Aaron L Albuck et al.
American journal of physiology. Heart and circulatory physiology, 320(2), H630-H641 (2020-11-10)
Peroxynitrite (PN), generated from the reaction of nitric oxide (NO) and superoxide, is implicated in the pathogenesis of ischemic and neurodegenerative brain injuries. Mitochondria produce NO from mitochondrial NO synthases and superoxide by the electron transport chain. Our objective was
Anupama Lakshmanan et al.
Nature protocols, 12(10), 2050-2080 (2017-09-08)
Gas vesicles (GVs) are a unique class of gas-filled protein nanostructures that are detectable at subnanomolar concentrations and whose physical properties allow them to serve as highly sensitive imaging agents for ultrasound and MRI. Here we provide a protocol for
Nadezhda S Sabeva et al.
Bio-protocol, 7(17) (2017-11-03)
We developed a protocol for photoconversion of endocytic marker FM1-43 followed by electron microscopy analysis of synaptic boutons at the Drosophila neuromuscular junction. This protocol allows detection of stained synaptic vesicle even when release rates are very low, such as
Yi Deng et al.
Analytica chimica acta, 868, 45-52 (2015-03-31)
Rapid and ultrasensitive detection of trace heavy metal mercury(II) ions (Hg(2+)) are of significant importance due to the induced serious risks for environment and human health. This presented article reports the gold nanoparticle-based dual labeling colorimetric method (Dual-COLO) for ultrasensitive
Karol Kadlec et al.
Talanta, 147, 44-49 (2015-11-26)
The novel approach for hydroxyapatite (HA) surface characterization was proposed. The main aim of this investigation was to estimate surface properties of HA as a biomaterial in real system i.e. in simulated body fluid (SBF). One of the method, which
William J Polacheck et al.
Nature protocols, 14(5), 1425-1454 (2019-04-07)
The vascular endothelium forms the inner lining of blood vessels and actively regulates vascular permeability in response to chemical and physical stimuli. Understanding the molecular pathways and mechanisms that regulate the permeability of blood vessels is of critical importance for
Christian Frezza et al.
Nature protocols, 2(2), 287-295 (2007-04-05)
Mitochondria participate in key metabolic reactions of the cell and regulate crucial signaling pathways including apoptosis. Although several approaches are available to study mitochondrial function in situ are available, investigating functional mitochondria that have been isolated from different tissues and
Anthony J Snyder et al.
Bio-protocol, 8(2) (2018-03-20)
Mammalian orthoreovirus (reovirus) utilizes pore forming peptides to penetrate host cell membranes. This step is essential for delivering its genome containing core particle during viral entry. This protocol describes an in vitro assay for measuring reovirus-induced pore formation.
Hsien-Sung Huang et al.
PloS one, 9(5), e98383-e98383 (2014-05-27)
Genomic imprinting describes an epigenetic process through which genes can be expressed in a parent-of-origin-specific manner. The monoallelic expression of imprinted genes renders them particularly susceptible to disease causing mutations. A large proportion of imprinted genes are expressed in the
Franziska E Uhl et al.
The European respiratory journal, 46(4), 1150-1166 (2015-05-02)
Chronic obstructive pulmonary disease (COPD) is characterised by a progressive loss of lung tissue. Inducing repair processes within the adult diseased lung is of major interest and Wnt/β-catenin signalling represents a promising target for lung repair. However, the translation of
Iva Stojkovska et al.
STAR protocols, 2(1), 100372-100372 (2021-03-19)
The accumulation of proteins into insoluble aggregates is a common feature of several neurodegenerative diseases. Aggregated α-synuclein is a major component of Lewy bodies that pathologically define Parkinson's disease (PD). Here, we present methods for the detection of pathogenic conformations
Erik Toraason et al.
Current biology : CB, 31(7), 1508-1514 (2021-03-20)
During meiosis, the maintenance of genome integrity is critical for generating viable haploid gametes.1 In meiotic prophase I, double-strand DNA breaks (DSBs) are induced and a subset of these DSBs are repaired as interhomolog crossovers to ensure proper chromosome segregation.
Xingmin Aaron Zhang et al.
Bio-protocol, 8(1) (2018-03-20)
Here we describe two assays to measure dense core vesicle (DCV) exocytosis-mediated cargo secretion in neuroendocrine cells. To conduct siRNA screens for novel genes in regulated DCV exocytosis, we developed a plate reader-based secretion assay using DCV cargo, NPY-Venus, and
Joshua E Mayfield et al.
Bio-protocol, 10(12), e3648-e3648 (2021-03-05)
Eukaryotic RNA polymerase II transcribes all protein-coding mRNAs and is highly regulated. A key mechanism directing RNA polymerase II and facilitating the co-transcriptional processing of mRNAs is the phosphorylation of its highly repetitive carboxyl-terminal domain (CTD) of its largest subunit
Shuai Jin et al.
Molecular cell, 79(5), 728-740 (2020-07-30)
Cytosine base editors (CBEs) generate C-to-T nucleotide substitutions in genomic target sites without inducing double-strand breaks. However, CBEs such as BE3 can cause genome-wide off-target changes via sgRNA-independent DNA deamination. By leveraging the orthogonal R-loops generated by SaCas9 nickase to
Abel Cuevas-Bermúdez et al.
Bio-protocol, 10(1), e3471-e3471 (2020-01-05)
We have adapted a previous procedure and improved an approach that we named yChEFs (yeast Chromatin Enriched Fractions) for purifying chromatin fractions. This methodology allows the easy, reproducible and scalable recovery of proteins associated with chromatin. By using yChEFs, we
Jason Yi et al.
Molecular biology of the cell, 27(22), 3591-3600 (2016-11-05)
Investigation of heterogeneous cellular structures using single-molecule localization microscopy has been limited by poorly defined localization accuracy and inadequate multiplexing capacity. Using fluorescent nanodiamonds as fiducial markers, we define and achieve localization precision required for single-molecule accuracy in dSTORM images.
Daniel J Tyrrell et al.
Journal of the American Heart Association, 9(23), e017820-e017820 (2020-11-24)
Background The blood-brain barrier (BBB) is critical for cerebrovascular health. Although aging impairs the integrity of the BBB, the mechanisms behind this phenomenon are not clear. As mitochondrial components activate inflammation as mitochondria become dysfunctional, we examined how aging impacts
Zuzana Nascakova et al.
International journal of molecular sciences, 22(7) (2021-05-01)
R-loops are three-stranded structures generated by annealing of nascent transcripts to the template DNA strand, leaving the non-template DNA strand exposed as a single-stranded loop. Although R-loops play important roles in physiological processes such as regulation of gene expression, mitochondrial
Jung Eun Kim et al.
Science advances, 7(10) (2021-03-05)
Stress is a key risk factor for dystonia, a debilitating motor disorder characterized by cocontractions of muscles leading to abnormal body posture. While the serotonin (5HT) system is known to control emotional responses to stress, its role in dystonia remains
Carola Städele et al.
Bio-protocol, 7(5), e2151-e2151 (2017-03-05)
This is a detailed protocol explaining how to perform extracellular axon stimulations as described in Städele and Stein, 2016. The ability to stimulate and record action potentials is essential to electrophysiological examinations of neuronal function. Extracellular stimulation of axons traveling
Niina Tani et al.
Bioorganic & medicinal chemistry, 22(23), 6655-6664 (2014-12-03)
Inhibition of CYP2A6-mediated nicotine metabolism can reduce cigarette smoking. We sought potent and selective CYP2A6 inhibitors to be used as leads for drugs useful in smoking reduction therapy, by evaluating CYP2A6 inhibitory effect of novel formyl, alkyl amine or carbonitrile
Maciej J Zelazowski et al.
Cell, 171(3), 601-614 (2017-09-26)
Faithful chromosome segregation in meiosis requires crossover (CO) recombination, which is regulated to ensure at least one CO per homolog pair. We investigate the failure to ensure COs in juvenile male mice. By monitoring recombination genome-wide using cytological assays and
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