細胞の文化<ol><li>ヒト胚性腎臓293（HEK293）細胞の保存培養は抗生物質のカクテル（ペニシリンのmlあたり100個および100μgを添加したダルベッコ改変イーグル培地（DMEM）で構成される&quot;中&quot;、で10cmの培養皿で増殖させるストレプトマイシン硫酸のml当たり）及び10％ウシ胎児血清（FBS）。 </li><li>実験用セルを設定するには、培地を吸引し、細胞をリン酸5mlで2回洗浄している生理食塩水（PBS）ソリューションをバッファ。 </li><li> PBSを除去し、0.25％トリプシンを含む溶液0.6 mLで置き換えられます。 </li><li>細胞がデタッチし始めているまで、皿を37˚Cで約2分間インキュベートする。 </li><li>培地5.5 mlのトリプシンを中和するために添加し、細胞を、血球計でカウントされます。 </li><li>細胞を、1ml当たり32万細胞に希釈し、ポリリジンコートした96穴培養プレートにウェルあたり0.1ミリリットルで分配される。 </li><li> 37˚C 8.8％CO2を含む大気中で組織培養インキュベーター中に皿を置き、24時間のために成長する。 </li></ol>トランスフェクション<ol><li>トランスフェクションのための準備では、実験条件とどのようなプラスミドの量はトランスフェクションされるごとに実行するかを多くの複製を考慮する必要があります。我々は日常的に三連のすべての条件を実行し、ウェルあたりのプラスミドDNAの50〜100 ngの合計をト​​ランスフェ。プラスミドミックスは、少なくとも記者はホタルルシフェラーゼと構成的プロモーターからの制御を発現するプラスミドウミシイタケルシフェラーゼを発現するコンストラクト含める必要があります。 </li><li>プラスミド溶液は1.5 mlのマイクロ遠心チューブにピペットと無血清および抗生物質を含まないDMEMのサンプルあたり10μLで補足されています。たとえば、20の井戸がそれぞれが50μgのDNAをトランスフェクトされる場合は、1μgのDNAに200μlのDMEMを加える。 </li><li> DMEM / DNA溶液に、μgのDNA当り3μlの&quot;トランジット293試薬&quot;（Mirus）を追加します。 ＆上下にピペッティングして混合し、少なくとも10分間室温で試料を放置する。 </li><li>この期間中、96ウェル培養プレートから培地を除去し、新鮮な培地Aの90μlのと交換してください</li><li>それぞれによくDMEM / DNA /トランジット293混合物の10μlを加える。 </li><li> 37˚C 8.8％CO2を含む大気中で組織培養インキュベーター中に皿を置き、24時間のために成長する。 </li></ol>貪食<ol><li>培地B（DMEMとHamのF12培地に加えて抗生物質の1:1混合物と10％FBS）を加えたml当たりゼロから1 mgで0.75μmのラテックスビーズを含む懸濁液を準備する。 </li><li> 96ウェル培養プレートからメディアを取り出して、培地Bのビーズ懸濁液0.1 mlを交換してください</li><li> 1000 × gで2分間プレートを回転させる</li><li>次に、細胞を6〜16時間、37˚Cで培養されています。条件とルシフェラーゼを駆動するために使用されるプロモーターに依存し、増加したレポーター遺伝子の発現は、1〜3時間後に検出することができます。 </li><li>同じケースではそれが30〜60分間ビーズで細胞をインキュベートするために必要となる場合があります、その後、PBSで2回洗浄には取り込まれなかったビーズを除去し、時間の追加の期間の細胞をインキュベートする前に新鮮な培地を追加することです。 </li></ol>デュアルGloTMルシフェラーゼアッセイ<ol><li>注：このキットを使用するための我々のプロトコルは、メーカーが推奨することからいくつかの点で異なります。これは、サンプルごとに必要な試薬の量を減らすために変更されています。 </li><li> -20˚Cで複数の小分けや店舗に1 M DTT、除算の溶液を調製します</li><li>の20mM Tris - HCl（pH7.8）中、10％（v / v）グリセロール、および0.5％（v / v）のトリトンX - 100を含む溶解バッファーを準備します。使用前に、一定量の実験に必要な量をと1xを0.5の最終濃度に1 M DTTを加えたプロテアーゼ阻害剤カクテルのmlあたり1μlを追加。残りのDTT溶液を再使用しないでください。 </li><li>初めてデュアルGloTMキットを使用する場合は、デュアルGloTMルシフェラーゼ基質（提供の一瓶に（キットに付属）デュアルGloTMルシフェラーゼバッファの一瓶の内容全体を転送することにより（ホタル）ルシフェラーゼ試薬を準備するキット付き）。 -20˚Cで10 mlのアリコートとストアに使用されていないルシフェラーゼ試薬を分ける</li><li>組織培養インキュベーターから96ウェルプレートを取り外し、氷の上に置きます。吸引によってメディアを削除する、40μlをウェルあたりの溶解バッファーを追加して、氷上で30分間プレートを残す。 </li><li>白、不透明な96ウェルプレート（OptiplateTM - 96、パーキンエルマーカタログ番号6005290）のアリコート15μlをウェルあたりのルシフェラーゼ試薬、およびその後細胞溶解液の15μLを追加で。 </li><li>室温で10分間プレートをインキュベートし、マイクロプレートリーダーで発光をお読みください。 </li><li>使用直前に、1部デュアルGloTMストップ＆グロを追加することにより、ウミシイタケルシフェラーゼの基質溶液の適当量を準備®、100部に試薬（キットに付属）デュアルGloTMストップ＆グロ®バッファー（キットに付属）。 </li><li>それぞれによく、10分間室温でインキュベートウミシイタケルシフェラーゼの基質溶液15μlのウェルあたりを、追加してから、再度発光を測定します。 </li></ol>

The authors have nothing to disclose.

studying membrane biogenesis with a luciferase-based reporter gene assay

膜の生合成中に別の脂質合成経路の調整を検討するためには、膜の生合成は、迅速かつ誘導的に発生した実験システムで動作するように便利です。我々は、細胞が急速に粒子の貪食時の材料のラッピングとして失わ膜プールを補充するために膜脂質を合成としてラテックスビーズの貪食は、これらの目的のために実用的であることを見出した。ここでは、プロメガからデュアルグロルシフェラーゼアッセイシステムを用いてルシフェラーゼベースのレポーター遺伝子アプローチと貪食誘導性遺伝子発現の変化を研究するための手順について説明します。

Watch this Scientific Journal Video about Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay at JoVE.com

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

ここで、我々はプロメガからデュアルGloTMルシフェラーゼアッセイシステムを用いてルシフェラーゼベースのレポーター遺伝子のアプローチで貪食誘導性遺伝子発現の変化を研究するための手順について説明します。

ルシフェラーゼベースのレポーター遺伝子アッセイで膜の生合成を研究して

To study the coordination of different lipid synthesis pathways during membrane biogenesis it is useful to work with an experimental system where membrane ...

studying-membrane-biogenesis-with-luciferase-based-reporter-gene

Research

JoVE Journal

Biology

12.8K ビュー.  Harvard. ここで、我々はプロメガからデュアルGloTMルシフェラーゼアッセイシステムを用いてルシフェラーゼベースのレポーター遺伝子のアプローチで貪食誘導性遺伝子発現の変化を研究するための手順について説明します。

ビデオ: Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

A Microfluidic Device with Groove Patterns for Studying Cellular Behavior

We describe a microfluidic device with microgrooved patterns for studying cellular behavior. This microfluidic platform consists of a top fluidic channel and a bottom microgrooved substrate. To fabricate the microgrooved channels, a top poly(dimethylsiloxane) (PDMS) mold containing the impression of the microfluidic channels was aligned and bonded to a microgrooved substrate. Using this device, mouse fibroblast cells were immobilized and patterned within microgrooved substrates (25, 50, 75, and 100 &mu;m wide). To study apoptosis in a microfluidic device, media containing hydrogen peroxide, Annexin V, and propidium iodide was perfused into the fluidic channel for 2 hours. We found that cells exposed to the oxidative stress became apoptotic. These apoptotic cells were confirmed by Annexin V that bound to phosphatidylserine at the outer leaflet of the plasma membrane during the apoptosis process. Using this microfluidic device with microgrooved patterns, the apoptosis process was observed in real-time and analyzed by using an inverted microscope containing an incubation chamber (37&deg;C, 5% CO2). Therefore, this microfluidic device incorporated with microgrooved substrates could be useful for studying the cellular behavior and performing high-throughput drug screening.

We describe a protocol for the fabrication of microfluidic devices that can enable cell capture and culture. In this approach patterned microstructures such as grooves within microfluidic channels are used to create low shear stress regions within which cell can dock.

細胞挙動を研究するための溝パターンを用いたマイクロ流体デバイス

We describe a microfluidic device with microgrooved patterns for studying cellular behavior.  This microfluidic platform consists of a top fluidic channel ...

生物学

Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells. We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions. We will demonstrate how to run several samples through a range of electroporation conditions so that an experiment can be conducted at the same time as optimization is performed. We will also show how optimal conditions identified using 96-well electroporation plates can be used with standard electroporation cuvettes, facilitating the switch from electroporation plates to electroporation cuvettes while maintaining the same electroporation efficiency. In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.

This procedure shows how to use the Gene Pulser MXcell electroporation system to rapidly and easily identify the best electroporation conditions for mouse embryonic fibroblasts (MEFs) or other primary cells. Considerations for troubleshooting are also discussed in the associated video.

高効率で初代培養細胞をトランスフェクトするためにジーンパルサーMXcellのエレクトロポレーションシステムの使用

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::&beta;-glucuronidase Reporter Gene Expression

The ability to introduce foreign genes into an organism is the foundation for modern biology and biotechnology. In the model flowering plant Arabidopsis thaliana, the floral-dip transformation method1-2 has replaced all previous methods because of its simplicity, efficiency, and low cost. Specifically, shoots of young flowering Arabidopsis plants are dipped in a solution of Agrobacterium tumefaciens carrying specific plasmid constructs. After dipping, the plants are returned to normal growth and yield seeds, a small percentage of which are transformed with the foreign gene and can be selected for on medium containing antibiotics. This floral-dip method significantly facilitated Arabidopsis research and contributed greatly to our understanding of plant gene function. In this study, we use the floral-dip method to transform a reporter gene, &beta;-glucuronidase (GUS), under the control of TSO2 promoter. TSO2, coding for the Ribonucleotide Reductase (RNR) small subunit3, is a cell cycle regulated gene essential for dNDP biosynthesis in the S-phase of the cell cycle. Examination of GUS expression in transgenic Arabidopsis seedlings shows that TSO2 is expressed in actively dividing tissues. The reported experimental method and materials can be easily adapted not only for research but also for education at high school and college levels.

Floral-dip Transformation of Arabidopsis thaliana to Examine pTSO2::&amp;#946;-glucuronidase Reporter Gene Expression

This article illustrates the floral-dip method of Agrobacterium tumefaciens -mediated transformation of Arabidopsis thaliana. By introducing a cell-cycle regulated promoter-reporter, pTSO2::&beta;-glucuronidase (GUS), into Arabidopsis, we illustrates how one detects GUS reporter expression in transgenic seedlings.

フローラルディップ変換のシロイヌナズナ調べに pTSO2：：β-グルクロニダーゼレポーター遺伝子発現

The ability to introduce foreign genes into an organism is the foundation for modern biology and biotechnology. In the model flowering plant Arabidopsis ...

Agrobacterium-Mediated Virus-Induced Gene Silencing Assay In Cotton

Cotton (Gossypium hirsutum) is one of the most important crops worldwide. Considerable efforts have been made on molecular breeding of new varieties. The large-scale gene functional analysis in cotton has been lagged behind most of the modern plant species, likely due to its large size of genome, gene duplication and polyploidy, long growth cycle and recalcitrance to genetic transformation1. To facilitate high throughput functional genetic/genomic study in cotton, we attempt to develop rapid and efficient transient assays to assess cotton gene functions.
Virus-Induced Gene Silencing (VIGS) is a powerful technique that was developed based on the host Post-Transcriptional Gene Silencing (PTGS) to repress viral proliferation2,3. Agrobacterium-mediated VIGS has been successfully applied in a wide range of dicots species such as Solanaceae, Arabidopsis and legume species, and monocots species including barley, wheat and maize, for various functional genomic studies3,4. As this rapid and efficient approach avoids plant transformation and overcomes functional redundancy, it is particularly attractive and suitable for functional genomic study in crop species like cotton not amenable for transformation.
In this study, we report the detailed protocol of Agrobacterium-mediated VIGS system in cotton. Among the several viral VIGS vectors, the tobacco rattle virus (TRV) invades a wide range of hosts and is able to spread vigorously throughout the entire plant yet produce mild symptoms on the hosts5. To monitor the silencing efficiency, GrCLA1, a homolog gene of Arabidopsis Cloroplastos alterados 1 gene (AtCLA1) in cotton, has been cloned and inserted into the VIGS binary vector pYL156. CLA1 gene is involved in chloroplast development6, and previous studies have shown that loss-of-function of AtCLA1 resulted in an albino phenotype on true leaves7, providing an excellent visual marker for silencing efficiency. At approximately two weeks post Agrobacterium infiltration, the albino phenotype started to appear on the true leaves, with 100% silencing efficiency in all replicated experiments. The silencing of endogenous gene expression was also confirmed by RT-PCR analysis. Significantly, silencing could potently occur in all the cultivars we tested, including various commercially grown varieties in Texas. This rapid and efficient Agrobacterium-mediated VIGS assay provides a very powerful tool for rapid large-scale analysis of gene functions at genome-wide level in cotton.

We present the detailed protocol for Agrobacterium-mediated virus-induced gene silencing (VIGS) assay in cotton. The tobacco rattle virus (TRV)-derived VIGS vectors were deployed to induce RNA silencing of cotton GrCLA1, Cloroplastos alterados 1 gene. The albino phenotype caused by silencing GrCLA1 was observed at the seedling stage within 2 weeks after inoculation.

コットンでアグロバクテリウムを介したウイルス誘発遺伝子サイレンシングアッセイ

Cotton (Gossypium hirsutum) is one of the most important crops worldwide. Considerable efforts have been made on molecular breeding of new varieties.  The ...

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3'UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

MicroRNAs (miRNAs) are important regulators of gene expression and play a role in many biological processes. More than 700 human miRNAs have been identified so far with each having up to hundreds of unique target mRNAs. Computational tools, expression and proteomics assays, and chromatin-immunoprecipitation-based techniques provide important clues for identifying mRNAs that are direct targets of a particular miRNA. In addition, 3'UTR-reporter assays have become an important component of thorough miRNA target studies because they provide functional evidence for and quantitate the effects of specific miRNA-3'UTR interactions in a cell-based system. To enable more researchers to leverage 3'UTR-reporter assays and to support the scale-up of such assays to high-throughput levels, we have created a genome-wide collection of human 3'UTR luciferase reporters in the highly-optimized LightSwitch Luciferase Assay System. The system also includes synthetic miRNA target reporter constructs for use as positive controls, various endogenous 3'UTR reporter constructs, and a series of standardized experimental protocols.
Here we describe a method for co-transfection of individual 3'UTR-reporter constructs along with a miRNA mimic that is efficient, reproducible, and amenable to high-throughput analysis.

Identifying Targets of Human microRNAs with the LightSwitch Luciferase Assay System using 3&#039;UTR-reporter Constructs and a microRNA Mimic in Adherent Cells

MicroRNAs (miRNAs) are important regulators of gene expression and have been shown to play a role in numerous biological processes. To better understand miRNA-UTR interactions, we have created a genome-wide collection of 3 UTR luciferase reporters paired with a novel luciferase gene and assay reagent, the LightSwitch system.

3&#39;UTR -レポーターコンストラクトとマイクロRNAを使用してLightSwitchルシフェラーゼアッセイシステムによるヒトマイクロRNAの識別対象は付着細胞へのミミック

MicroRNAs (miRNAs) are important regulators of gene expression and play a role in many biological processes.  More than 700 human miRNAs have been ...

A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System

Glucocorticoid stress hormones and their artificial derivatives are widely used drugs to treat inflammation, but long-term treatment with glucocorticoids can lead to severe side effects. Test systems are needed to search for novel compounds influencing glucocorticoid signaling in vivo or to determine unwanted effects of compounds on the glucocorticoid signaling pathway. We have established a transgenic zebrafish assay which allows the measurement of glucocorticoid signaling activity in vivo and in real-time, the GRIZLY assay (Glucocorticoid Responsive In vivo Zebrafish Luciferase activitY). The luciferase-based assay detects effects on glucocorticoid signaling with high sensitivity and specificity, including effects by compounds that require metabolization or affect endogenous glucocorticoid production. We present here a detailed protocol for conducting chemical screens with this assay. We describe data acquisition, normalization, and analysis, placing a focus on quality control and data visualization. The assay provides a simple, time-resolved, and quantitative readout. It can be operated as a stand-alone platform, but is also easily integrated into high-throughput screening workflows. It furthermore allows for many applications beyond chemical screening, such as environmental monitoring of endocrine disruptors or stress research.

We describe the procedure and data analysis of a chemical screening system for glucocorticoid stress hormone signaling using zebrafish larvae: the Glucocorticoid Responsive In vivo Zebrafish Luciferase activitY (GRIZLY) assay. The assay sensitively and specifically detects effects on glucocorticoid signaling by compounds that require metabolization or affect endogenous glucocorticoid production.

ゼブラフィッシュ幼虫ルシフェラーゼレポーターシステムを用いたグルココルチコイドシグナル伝達のための化学スクリーニング法

Glucocorticoid stress hormones and their artificial derivatives are widely used drugs to treat inflammation, but long-term treatment with glucocorticoids ...

Reporter-based Growth Assay for Systematic Analysis of Protein Degradation

Protein degradation by the ubiquitin-proteasome system (UPS) is a major regulatory mechanism for protein homeostasis in all eukaryotes. The standard approach to determining intracellular protein degradation relies on biochemical assays for following the kinetics of protein decline. Such methods are often laborious and time consuming and therefore not amenable to experiments aimed at assessing multiple substrates and degradation conditions. As an alternative, cell growth-based assays have been developed, that are, in their conventional format, end-point assays that cannot quantitatively determine relative changes in protein levels.
Here we describe a method that faithfully determines changes in protein degradation rates by coupling them to yeast cell-growth kinetics. The method is based on an established selection system where uracil auxotrophy of URA3-deleted yeast cells is rescued by an exogenously expressed reporter protein, comprised of a fusion between the essential URA3 gene and a degradation determinant (degron). The reporter protein is designed so that its synthesis rate is constant whilst its degradation rate is determined by the degron. As cell growth in uracil-deficient medium is proportional to the relative levels of Ura3, growth kinetics are entirely dependent on the reporter protein degradation.
This method accurately measures changes in intracellular protein degradation kinetics. It was applied to: (a) Assessing the relative contribution of known ubiquitin-conjugating factors to proteolysis (b) E2 conjugating enzyme structure-function analyses (c) Identification and characterization of novel degrons. Application of the degron-URA3-based system transcends the protein degradation field, as it can also be adapted to monitoring changes of protein levels associated with functions of other cellular pathways.

Here we describe a robust biological assay for quantifying the relative rate of proteolysis by the ubiquitin-proteasome system. The assay readout is yeast growth rate in liquid culture, which is dependent on the cellular levels of a reporter protein comprising a degradation signal fused to an essential metabolic marker.

タンパク質分解の体系的な分析のためのレポーターベースの成長アッセイ

Protein degradation by the ubiquitin-proteasome system (UPS) is a major regulatory mechanism for protein homeostasis in all eukaryotes. The standard ...

Transfecting RAW264.7 Cells with a Luciferase Reporter Gene

Transfection of desired genetic materials into cells is an inevitable procedure in biomedical research studies. While numerous methods have been described, certain types of cells are resistant to many of these methods and yield low transfection efficiency1, potentially hindering research in those cell types. In this protocol, we present an optimized transfection procedure to introduce luciferase reporter genes as a plasmid DNA into the RAW264.7 macrophage cell line. Two different types of transfection reagents (lipid-based and polyamine-based) are described, and important notes are given throughout the protocol to ensure that the RAW264.7 cells are minimally altered by the transfection procedure and any experimental data obtained are the direct results of the experimental treatment. While transfection efficiency may not be higher compared to other transfection methods, the described procedure is robust enough for detecting luciferase signal in RAW264.7 without changing the physiological response of the cells to stimuli.

Transfection into the macrophage cell line, RAW264.7, is difficult due to the cell&#8217;s natural response against foreign materials. We described here a gentle yet robust procedure for transfecting luciferase reporter genes into RAW264.7 cells.

ルシフェラーゼレポーター遺伝子でRAW264.7細胞をトランスフェクト

Transfection of desired genetic materials into cells is an inevitable procedure in biomedical research studies. While numerous methods have been ...

Visualizing Stromule Frequency with Fluorescence Microscopy

Stromules, or "stroma-filled tubules", are narrow, tubular extensions from the surface of the chloroplast that are universally observed in plant cells but whose functions remain mysterious. Alongside growing attention on the role of chloroplasts in coordinating plant responses to stress, interest in stromules and their relationship to chloroplast signaling dynamics has increased in recent years, aided by advances in fluorescence microscopy and protein fluorophores that allow for rapid, accurate visualization of stromule dynamics. Here, we provide detailed protocols to assay stromule frequency in the epidermal chloroplasts of Nicotiana benthamiana, an excellent model system for investigating chloroplast stromule biology. We also provide methods for visualizing chloroplast stromules in vitro by extracting chloroplasts from leaves. Finally, we outline sampling strategies and statistical approaches to analyze differences in stromule frequencies in response to stimuli, such as environmental stress, chemical treatments, or gene silencing. Researchers can use these protocols as a starting point to develop new methods for innovative experiments to explore how and why chloroplasts make stromules.

Protocols to investigate the dynamics of chloroplast stromules, the stroma-filled tubules that extend from the surface of chloroplasts, are described.

蛍光顕微鏡でストロミュール頻度を可視化

Stromules, or "stroma-filled tubules", are narrow, tubular extensions from the surface of the chloroplast that are universally observed in plant cells but ...

An In Vitro Model for Studying Cellular Transformation by Kaposi Sarcoma Herpesvirus

Kaposi sarcoma (KS) is an unusual tumor composed of proliferating spindle cells that is initiated by infection of endothelial cells (EC) with KSHV, and develops most often in the setting of immunosuppression. Despite decades of research, optimal treatment of KS remains poorly defined and clinical outcomes are especially unfavorable in resource-limited settings. KS lesions are driven by pathological angiogenesis, chronic inflammation, and oncogenesis, and various in vitro cell culture models have been developed to study these processes. KS arises from KSHV-infected cells of endothelial origin, so EC-lineage cells provide the most appropriate in vitro surrogates of the spindle cell precursor. However, because EC have a limited in vitro lifespan, and as the oncogenic mechanisms employed by KSHV are less efficient than those of other tumorigenic viruses, it has been difficult to assess the processes of transformation in primary or telomerase-immortalized EC. Therefore, a novel EC-based culture model was developed that readily supports transformation following infection with KSHV. Ectopic expression of the E6 and E7 genes of human papillomavirus type 16 allows for extended culture of age- and passage-matched mock- and KSHV-infected EC and supports the development of a truly transformed (i.e., tumorigenic) phenotype in infected cell cultures. This tractable and highly reproducible model of KS has facilitated the discovery of several essential signaling pathways with high potential for translation into clinical settings.

An In Vitro Model for Studying Cellular Transformation by Kaposi Sarcoma Herpesvirus

Kaposi sarcoma (KS) is a tumor induced by infection with the oncogenic virus human herpesvirus-8/KS herpesvirus (HHV-8/KSHV). The endothelial cell culture model described here is uniquely suited for studying the mechanisms by which KSHV transforms host cells.

カポジ肉腫ヘルペス ウイルスによる細胞変換を勉強するための in Vitroモデル

Kaposi sarcoma (KS) is an unusual tumor composed of proliferating spindle cells that is initiated by infection of endothelial cells (EC) with KSHV, and ...