Результаты поиска

Hetero-oligomerization of the P2Y11 receptor with the P2Y1 receptor controls the internalization and ligand selectivity of the P2Y11 receptor.

Nucleotides signal through purinergic receptors such as the P2 receptors, which are subdivided into the ionotropic P2X receptors and the metabotropic P2Y receptors. The diversity of functions within the purinergic receptor family is required for the tissue-specificity of nucleotide signalling. In the present study, hetero-oligomerization between two metabotropic P2Y receptor subtypes is established. These receptors, P2Y1 and P2Y11, were found to associate together when co-expressed in HEK293 cells. This association was detected by co-pull-down, immunoprecipitation and FRET (fluorescence resona…

Rapid measurements of intracellular calcium using a fluorescence plate reader.

Intracellular calcium is a universal second messenger that can serve as a broad-based measure of receptor activity. Recent developments in m … re able to detect and quantify activity from muscarinic and beta-adrenergic receptors endogenous to HEK293 cells and detect calcium signals generated by activation of Gi-coupled recombinant mu-opioid and dopamine D2L receptors, and the Gs-coupled melanocortin subtype 4 (MC4) receptor. Fluorescence signals, stable in HEK293 cells, required the use of Oregon Gre …

Nuclear translocation of STAT5 initiates iron overload in huntington's disease by up-regulating IRP1 expression.

Mutant huntingtin (mHtt) proteins interact to form aggregates, disrupting cellular functions including transcriptional dysregulation and iron imbalance in patients with Huntington's disease (HD) and mouse disease models. Previous studies have indicated that mHtt may lead to abnormal iron homeostasis by upregulating the expression of iron response protein 1 (IRP1) in the striatum and cortex of N171-82Q HD transgenic mice, as well as in HEK293 cells expressing the N-terminal fragment of mHtt containing 160 CAG repeats. However, the mechanism underlying the upregulation of IRP1 remains unclear. W…

Impact of cryopreservation on immune cell subset frequencies in human peripheral blood mononuclear cells following assessment of monoclonal antibodies to CD39 and P2X7.

Peripheral blood mononuclear cells (PBMCs) are commonly cryopreserved for immune cell profiling analyses, like flow cytometry. Cryopreservat … bound human embryonic kidney (HEK) 293 cells transfected with human CD39, but not mock-transfected HEK293 cells. In contrast, an anti-human P2X7 mAb (clone 1058613) bound similarly to HEK293 cells transfected with human P2X7 and non-transfected HEK293 cells. Using flow cytometry, this study then assessed the impact of cryopreservation on the population freque …

Differentiation between human ClC-2 and CFTR Cl- channels with pharmacological agents.

It has been difficult to separate/identify the roles of ClC-2 and CFTR in Cl(-) transport studies. Using pharmacological agents, we aimed to … -dependent manner equipotently. DASU-02 inhibited forskolin-IBMX-stimulated Cl(-) currents in hCFTR/HEK293 cells, but not lubiprostone-stimulated Cl(-) currents in hClC-2/293EBNA cells. In T84 cells with basolateral nystatin or 1-ethyl-2-benzimidazolinone (1-EBIO), lubiprostone-stimulated and forskolin-IBMX-cyclosporin A (FICA)-stimulated Isc components were o …

Functional Validation of a Novel <italic>PBX1</italic> Missense Variant in a 46,XY Girl.

INTRODUCTION: The pre-B-cell leukemia transcription factor encoded by PBX1 is expressed throughout human embryonic stages. Accumulating case … T reduced protein stability and hampered nuclear translocation of PBX1 in the inducible Flp-In TREx HEK293 cells. Induction with tetracycline significantly decreased cell proliferation in both Flp-In HEK293 PBX1 WT and p.R237T cells compared to untransfected HEK293 cells, while adhesive ability was not different. RNA sequencing identified differentially expres …

The chemopreventive effects of aged garlic extract against cadmium-induced toxicity.

Garlic has been reported in many previous studies as a potent chemopreventive agent. The protective effect of garlic has been ascribed to the presence of organosulphur compounds (OSC). In this study, the efficacy of aged garlic extract (AGE) compared to diallyl disulfide (DADS) in protecting against toxicity induced by cadmium (Cd) in 1321N1 and HEK293 cells was investigated. The involvement of the transcription factor Nrf2 in this protection was also examined. The results show that AGE significantly prevented loss of cell viability in Cd-treated 1321N1 and HEK293 cells. In comparison DADS had…

Evaluation of cytotoxicity attributed to thimerosal on murine and human kidney cells.

Renal inner medullary collecting duct cells (mIMCD3) and human embryonic kidney cells (HEK293) were used for cytoscreening of thimerosal and mercury chloride (HgCl2). Thimerosal and HgCl2 acted in a concentration-dependent manner. In mIMCD3 cells the 24-h LC50 values for thimerosal, thiosalicylic acid, 2,2-dithiosalicylic acid, and 2-sulfobenzoic acid were 2.9, 2200, >1000, and >10,000 microM, respectively. The 24-h LC50 value for HgCl2 in mIMCD3 cells was 40 microM. In HEK293 cells, the 24-h LC50 value for thimerosal was 9.5 microM. These data demonstrate that the higher cytotoxicity produced…

OLFM4-RET fusion is an oncogenic driver in small intestine adenocarcinoma.

Small intestine adenocarcinoma is a rare intestinal malignancy with distinct clinical, pathological, and molecular characteristics. Recently … r grade and advanced tumor stage. Expression of OLFM4-RET fusion-induced cellular transformation in HEK293 cells and blocked RET-induced inhibition of colony growth in HuTu 80 small intestine adenocarcinoma cells. Further, expression of OLFM4-RET activated the RAS-RAF-MAPK and STAT3 cell signaling pathways in both HEK293 cells and HuTu 80 cells. OLFM4-RET expr …

Development and Application of a Cell-Based Assay for LRP4 Antibody Associated With Myasthenia Gravis.

BACKGROUND AND PURPOSE: Among patients with double-seronegative myasthenia gravis (dSN-MG) who do not have detectable antibodies against ace … was transfected into human embryonic kidney 293 (HEK293) cells. LRP4 expression in the transfected HEK293 cells was assessed using the reverse-transcription polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemistry. The CBA included 252 sera collected from 202 patients with MG and 38 with other neuromuscular diseases, and 12 healthy contro …

A luciferase complementation assay system using transferable mouse artificial chromosomes to monitor protein-protein interactions mediated by G protein-coupled receptors.

G protein-coupled receptors (GPCRs) are seven-transmembrane domain receptors that interact with the β-arrestin family, particularly β-arrest … ely, via the sequential integration of multiple vectors (SIM) system. Each MAC was transferred into HEK293 cells by microcell-mediated chromosome transfer (MMCT). LCAs using the established HEK293 cell lines resulted in 35,000 photon counts upon somatostatin stimulation for ARRB1-MAC4 with transient transfection of the somatostatin receptor 2 (SSTR2) expressio …

Nicastrin haploinsufficiency alters expression of type I interferon-stimulated genes: the relationship to familial hidradenitis suppurativa.

BACKGROUND: Hidradenitis suppurativa (HS), also called acne inversa, is a chronic skin disease. The symptoms can be severe, and include inte … nt increase in nuclear factor kappa B signalling in response to tumour necrosis factor treatment in HEK293 cells genome-edited for reduced NCSTN. CONCLUSIONS: Our data suggest a role for increased keratinocyte inflammatory responsiveness in familial HS. Confirming this phenotype and characterizing additional effects in different cell types will require study …

The survival motor neuron protein interacts with the transactivator FUSE binding protein from human fetal brain.

To identify interacting proteins of survival motor neuron (SMN) in neurons, a fetal human brain cDNA library was screened using the yeast two-hybrid system. One identified group of SMN interacting clones encoded the DNA transactivator FUSE binding protein (FBP). FBP overexpressed in HEK293 cells or endogenously expressed in fetal and adult mouse brain bound specifically in vitro to recombinant SMN protein. Furthermore, an anti-FBP antibody specifically co-immunoprecipitated SMN when both proteins were overexpressed in HEK293 cells. These results demonstrate that FBP is a novel interacting part…

Development of a perfusion process for serum-free adenovirus vector herpes zoster vaccine production.

Herpes zoster is caused by reactivation of the varicella zoster virus (VZV). Researching and developing a herpes zoster vaccine will help to decrease the incidence of herpes zoster. To increase the bioreactor productivity, a serum-free HEK293 cell perfusion process with adenovirus vector herpes zoster (rAd-HZ) vaccine production was developed efficiently using the design of experiment (DoE) method. First, serum-free media for HEK293 cells were screened in both batch and semi-perfusion culture modes. Then, three optimal media were employed in a medium mixture design to improve cell culture perf…

Screening of plant extracts for human tyrosinase inhibiting effects.

Screening for tyrosinase (TYR) inhibitors potentially useful for control of skin pigmentation has been hampered by the limited availability of human TYR. To overcome this hurdle, we have established human embryonic kidney (HEK293)-TYR cells that constitutively express human TYR. In the current study, we assayed human TYR inhibition activities of 50 plant extracts using the lysates of transformed HEK293-TYR cells. The strongest inhibition of human TYR was shown by the extract of Vaccinium bracteatum Thunberg, followed by the extract of Morus bombycis Koidzumi. The former extract did not inhibit

Инженерная спящая красавица транспозон как эффективная система оптимизации производства шимпа Аденовира.

Спящая красавица (SB) — первый транспозон ДНК, используемый для эффективного транспонирования в позвоночных клетках, открывая новые приложения для генной инженерии и генной терапии. Система передачи генов на основе транспозона содержит благоприятные особенности невирусных векторов и привлекательный профиль безопасности. Здесь мы использовали SB для разработки клеток HEK293 для оптимизации производства шимпанзе Аденовектора (chAD), относящегося к человеческому мастаденовирусу C. На сегодняшний день переносчики chAD применяются в нескольких клинических условиях для инфекционных заболеваний, посл

Совпадение в аннотации оригинала
Sleeping Beauty (SB) is the first DNA transposon employed for efficient transposition in vertebrate cells, opening new applications for genetic engineering and gene therapies. A transposon-based gene delivery system holds the favourable features of non-viral vectors and an attractive safety profile. Here, we employed SB to engineer HEK293 cells for optimizin…

A proline-rich peptide improves cell transfection of solid lipid nanoparticle-based non-viral vectors.

The aim of this work was to improve the transfection efficacy of solid lipid nanoparticle (SLN)-based non-viral vectors into ARPE-19 cells t … The presence of SAP in the formulations did not affect the viability of HEK293 or ARPE-19 cells. In HEK293 cells, SAP enabled greater uptake of the vectors, and an SAP to DNA ratio of 50:1 was sufficient for enhancing transfection. In contrast, in ARPE-19 cells, SAP induced a change in the dominant entrance mechanism, from clathrin endocytosis to caveolae/raft …

Activating antibodies to the calcium-sensing receptor in two patients with autoimmune hypoparathyroidism.

Autoimmune hypoparathyroidism is thought to result from immune-mediated destruction of the parathyroid glands. We encountered two patients w … cells transfected with the extracellular calcium-sensing receptor (CaR) but not with nontransfected HEK293 cells. The antibodies also reacted with the same bands on Western analysis of extracts of bovine parathyroid tissue and CaR-transfected HEK293 cells that were identified by an authentic, polyclonal, anti-CaR antiserum and reacted with several peptides wit …

A novel tacrine-dihydropyridine hybrid (-)SCR1693 induces tau dephosphorylation and inhibits Aβ generation in cells.

AChE inhibitors are the first choice for the treatment of Alzheimer׳s disease (AD), but they could only delay the progression of cognitive a … in low concentrations such as 0.4, 2, 4μM for 24h did not affect the cell viability of HEK293/tau (HEK293 cells stably transfected with human tau40) and N2a/APP (N2a cells stably transfected with human APP) cells; (2) long-term treatment of cells with (-)SCR1693 (0.4, 2, 5μM) (24h) induced tau dephosphorylation and reduced the total tau level in HEK293/tau ce …

Evaluation of IRES-mediated, cell-type-specific cytotoxicity of poliovirus using a colorimetric cell proliferation assay.

PVS-RIPO is a recombinant oncolytic poliovirus designed for clinical application to target CD155 expressing malignant gliomas and other mali … , PVS-RIPO inhibited proliferation of U87-MG astrocytoma cells in a dose-dependent manner. However, HEK293 cells were much less susceptible to cell killing by PVS-RIPO. In contrast, the Sabin type 1 live attenuated poliovirus vaccine strain (PV(1)S) was cytotoxic to both HEK293 and U87-MG cells. The correlation between expression of CD155 and cytotoxicity was …