Name: in situ immunofluorescent staining of autophagy in muscle stem cells
Uploaded: 2017-06-12T15:00:00
Description: Watch this Scientific Journal Video about In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells at JoVE.com

Bu çalışma, NIAMS AR064873, Epigen Project PB tarafından desteklendi. P01.001.019 / Progetto Bandiera Epigenomica IFT - LL

Fareler, standart hayvan tesisi prosedürlerine göre yetiştirildi ve muhafaza edildi ve tüm deney protokolleri, Hayvan Refahı Güvencesi ve İtalyan Sağlık Bakanlığı&#39;na göre dahili Hayvan Araştırma Etik Kurulu tarafından onaylandı ve NIH&#39;nin Bakım ve Kullanım Kılavuzu&#39;na Laboratuar hayvanları. 1. Otofajik Flux&#39;ın Kası Yaralanması ve In Vivo Blokları <ol><li> Kas yaralanması. <ol><li> Akut iskelet kası hasarını uyandırmak ...

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	<li>Bentzinger, C. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+response+of+skeletal+muscles+to+mTORC1+signaling+during+atrophy+and+hypertrophy.">Differential response of skeletal muscles to mTORC1 signaling during atrophy and hypertrophy.</a> Skelet Muscle. 3 (1), (2013).</li><li>Chakkalakal, J. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+aged+niche+disrupts+muscle+stem+cell+quiescence.">The aged niche disrupts muscle stem cell quiescence.</a> Nature. 490 (7420), 355-360 (2012).</li><li>Jang, Y. 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<html> Bu protokol telafi edici kas rejenerasyonu sırasında iskelet kası kök hücrelerinde otofajinin nasıl izleneceğini açıklamaktadır. LC3 ve MyoD&#39;nin birlikte lekelenmesi için çeşitli antikorlar denendi ve fare dokusu kesitlerinde çalışan ve başarılı sonuçlar oluşturan antikorlar burada listelenmiştir (bkz. Malzeme Tablosu ). Başarılı bir boyama için metanol ile permeabilizasyon (bkz. Adım 3.2.2) önerilir. Bu protokolün sın...

İskelet kası rejenerasyonu, yetişkin kök hücreler (Kas Uydu Hücreleri, MuSCs) ve rejeneratif süreçte yer alan diğer hücre tipleri arasındaki etkileşimin bir sonucudur. Kasların homeostazı ve işlevselliği, kas nişinden ve sistemik ipuçlarından 1 , 2 oluşan kombine sinyaller tarafından sağlanır. Yaşam boyu MuSC işlevselliği, kas nişindeki değişiklikler ve sistemik ipuçları bildirildi ve yaşlılarda fonksiyonel kapasitelerin azalmasına neden oldu3. MuSC&#39;ler bazal tabakanın altında bir niş halinde bulunur ve kas hasarına bağlı olarak hasar görmüş kasları onarmak için harekete geçirilir 4 , 5 . Üretken bir rejeneratif tepki sağlamak için, MuSC&#39;lerin sessizliğe, kendi kendini yenilemeye ve proliferatif genişleme aşaması için gerekli olan farklı süreçleri koordine etmeleri çok önemlidirMiyojenik farklılaşma ile 6 . Yaşlılarda ve kas kronik hastalıklarında, bu işlevlerin tümüyle tehlikeye girer ve kas işlevlerinde değişikliklere yol açar 2 , 3 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 . Macroautophagy (bundan böyle otofaji olarak anılacaktır), doku homeostazını korumak için hayati bir biyolojik süreç olarak ortaya çıkmaktadır14. Otofajik süreç, sitoplazmanın, organellerin ve proteinlerin kısımlarının sonunda lizozom yolu ile bozunduğu toksik moleküllerin çıkarılmasını ve makromolefin geri dönüşümünü teşvik eden kaçakçılık mekanizmalarını kapsarcules. Bu, stres veya diğer olumsuz koşullar altında hücre ve doku adaptasyonunu desteklemek için enerji açısından zengin bileşikler sağlar 15 , 16 . Otofajy, hücre sağkalımı aktivitesi ile birlikte, hücre dokusu bağlamına ( örn. Kanser dokusuna karşı normal) ve stres uyarımının tipine ( 17 , 18 ) bağlı olarak bir hücre ölüm indüksiyonu olarak da çalışabilir. Yeni kanıtlar, kas kütlesi ve miyofiber bütünlüğünü korumak için otofajinin gerekli olduğunu göstermektedir 19,20 ve Duchenne Musküler Distrofi (DMD) dahil olmak üzere farklı kas distrofileri 21 , 22 , 23&#39;de bozulmuş olduğu bildirilmiştir 24 , 25 , 26 , 27 </suBenzer şekilde, kas kütlesi kaybına (sarkopenik olarak) 32 , 33 , 34 , 35 yaşlarında yaşlılarda otofajik progresifin ilerleyici bir azalması gözlemlenmiştir ( 32 , 33 , 34 , 35 ) , 35 , 36 , 37 ve myofiber sağkalımda 38 . Otofaji ile iskelet kasının rejeneratif potansiyeli arasındaki yakın ilişki Wagers laboratuarında yapılan bir çalışmada öngörüldü ve kalori kısıtlamasının MuSC&#39;nin kullanılabilirliğini ve aktivitesini arttırdığını gösterdi 39 . Bu hayırFoxo3-Notch ekseni, kendi kendine yenileme 40 sırasında otofajik işlemi aktive etti ve MuSC geçişi durdurma durumundan prolifere hale getirme durumuna geçtiğine dair son gözlem tarafından desteklendi 41 . Bu veriler, genç yaşlı ve geriatrik MuSC&#39;lere bazal otofajinin kademeli olarak azaltılması ve 42 yaşındaki MUHS&#39;lerin sayısal ve fonksiyonel düşüşüyle ​​birlikte kabul edilmektedir. Yeni bir makalede, otofaji ile DMD progresyonunun erken safhalarını ayıran telafi edici kas rejenerasyonu arasında yakın bir ilişki olduğunu gösterdik. Buna göre, kas yenilenmesinden ödün verildiğinde ve fibrotik doku birikimi oluştuğunda, hastalığın ilerlemesinin ilerleyen aşamalarında azaltılmış otofajik akı bulduk. İlginçtir ki, yenilenen koşullarda otofajinin MuSC&#39;lerde aktive edildiğini ve otofajik süreci modüle etmenin MuSC aktivasyonunu ve fu&#39;yu etkilediğini gösterdikNasyonellik 30 . Tamamı, bu veriler normal ve patolojik koşullarda ve ömrü boyunca kas yenilenmesi sırasında MuSC&#39;lerde otofajik süreci araştırmanın aciliyetini vurguluyor. Burada, kas yenileme koşullarında MuSC&#39;lerde otofajik işlemi izleyen bir protokol, otofajinin 43&#39;ü olan mikrotübüle bağlı protein 1A / 1B-hafif zincir 3 (LC3) ve MyoD&#39;nin bir markörü olan in situ immün boyama işlemi gerçekleştirerek Miyogenik soy, kontrol ve yaralı farelerin kas dokusu bölümlerinde. Bildirilen metodoloji, kas rejenerasyonunun düzenlenmesinde önemli bir rol oynayan MuSC adlı belirli bir hücre bölmesindeki otofajik sürecin izlenmesine izin verir.

<table border="0" cellpadding="0" cellspacing="0" width="906">
	<tbody>
		<tr height="19">
			<td height="19" style="height:19px;width:332px;">C57BL/6J</td>
			<td style="width:251px;">The Jackson Laboratory</td>
			<td style="width:128px;">000664</td>
			<td style="width:196px;">WT mice</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Cardiotoxin 1</td>
			<td>Latoxan</td>
			<td>L8102</td>
			<td></td>
		</tr>
		<tr height="32">
			<td height="32" style="height:32px;">Millex-VV</td>
			<td>Merck Millipore</td>
			<td>SLVV033RS</td>
			<td style="width:196px;">Syringe Filter Unit, 0.1 &#181;m, PVDF, 33 mm, gamma sterilized</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;">Chloroquine diphosphate salt</td>
			<td>Sigma-Aldrich</td>
			<td>C6628</td>
			<td style="width:196px;">Caution: 
			Harmful if swallowed</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;">BD Micro-Fine + 0,5 mL</td>
			<td>BD</td>
			<td>324825</td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Tissue-Tek O.C.T. compound</td>
			<td>Sakura Finetek</td>
			<td>25608-930</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Tissue-Tek Cryomold Intermediate</td>
			<td>Sakura Finetek</td>
			<td>4566</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">2-Methylbutane</td>
			<td>Sigma-Aldrich</td>
			<td>277258</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Hematoxylin Solution, Harris Modified</td>
			<td>Sigma-Aldrich</td>
			<td>HHS32</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Eosin Y solution, alcoholic</td>
			<td>Sigma-Aldrich</td>
			<td>HT110132</td>
			<td></td>
		</tr>
		<tr height="122">
			<td height="122" style="height:122px;">o-Xylene</td>
			<td>Sigma-Aldrich</td>
			<td>X1040</td>
			<td style="width:196px;">Caution: 
			Flammable liquid and vapour; May be fatal if swallowed and enters airways; Harmful in contact with skin; May cause respiratory irritation; Causes serious eye irritation</td>
		</tr>
		<tr height="136">
			<td height="136" style="height:136px;">Paraformaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>P6148</td>
			<td style="width:196px;">Caution: 
			Flammable solid; Harmful if swallowed; Causes skin irritation; May cause an allergic skin reaction; Causes serious eye damage; May cause respiratory irritation; Suspected of causing cancer</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">DPBS, no calcium, no magnesium</td>
			<td>Thermo Fisher Scientific</td>
			<td>14190-094</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Bovine Serum Albumin</td>
			<td>Sigma-Aldrich</td>
			<td>A7030</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Glycerol</td>
			<td>Sigma-Aldrich</td>
			<td>G5516</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:332px;">Eukitt - Quick-hardening mounting medium</td>
			<td>Sigma-Aldrich</td>
			<td>3989</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">AffiniPure Fab Fragment Goat Anti-Mouse IgG (H+L)</td>
			<td>Jackson ImmunoResearch</td>
			<td>115-007-003</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">LC3B Antibody</td>
			<td>Cell signaling Technology</td>
			<td>2775</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:332px;">Monoclonal mouse anti-MyoD 
			(concentrated) clone 5.8A</td>
			<td>DAKO - Agilent Pathology Solutions</td>
			<td>M3512</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Laminin-2 (&#945;-2-chain) monoclonal antibody</td>
			<td>Enzo Life Sciences</td>
			<td>4H8-2</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:332px;">Alexa Fluor 488 Goat Anti-Rabbit IgG (H+L)</td>
			<td style="width:251px;">Life technologies</td>
			<td style="width:128px;">A11008</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:332px;">Alexa Fluor 594 Goat Anti-Mouse IgG (H+L)</td>
			<td style="width:251px;">Life technologies</td>
			<td style="width:128px;">A11005</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:332px;">Alexa Fluor Goat Anti-Rat IgM Antibody</td>
			<td style="width:251px;">Life technologies</td>
			<td style="width:128px;">A21248</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:332px;">DAPI (4&#39;,6-Diamidino-2-Phenylindole, Dihydrochloride)</td>
			<td>Thermo Fisher Scientific</td>
			<td>D1306</td>
			<td></td>
		</tr>
	</tbody>
</table>

in situ immunofluorescent staining of autophagy in muscle stem cells

Artan kanıtlar, doku homeostazını korumak için önemli bir düzenleyici süreç olarak otofajiye işaret ediyor. Otofajinin iskelet kası gelişiminde ve yenilenmesinde rol aldığı bilinmektedir ve otofajik süreç birkaç kas patolojisinde ve yaşla ilgili kas bozukluklarında tanımlanmıştır. Otofajik işlemin kısa bir süre önce açıklanan, kas onarımında uydu hücrelerinin işlevsel tükenişiyle ilişkili olan bloğu, aktif otofajinin üretken kas rejenerasyonu ile birleştiğini desteklemektedir. Bu veriler, kas distrofileri gibi hem normal hem de patolojik koşullarda kas rejenerasyonu sırasında uydu hücresinin aktivasyonunda otofajinin önemli rolünü ortaya çıkarmaktadır. Burada, kas rejeneratif koşulları sırasında yetişkin Kas Hücresi (MuSC) bölmesindeki otofajik süreci izlemek için bir protokol sunuyoruz. Bu protokol LC3&#39;ün in situ immünofloresans görüntülemesini gerçekleştirmek için kurulum metodolojisini açıklamaktadır.Utofagy markeri ve MyoD, miyojenik soy işaretleyicisini, kontrol ve hasar gören farelerdeki kas dokusu kesitleri içinde göstermektedir. Bildirilen metodoloji, kas rejenerasyonunun düzenlenmesinde merkezi bir rol oynayan bir özel hücre bölmesindeki MuSC bölmesindeki otofajik prosedürün izlenmesine izin verir.

Bu protokol, kas rejenerasyonu sırasında MuSC&#39;lerde otofajiyi saptamak için etkin bir in situ metodu tarif eder. CTX In Vivo Tedaviler: TA kaslarındaki kas hasarını uyandırmak ve kontrolsüz kasları kontrol olarak kullanmak için CTX kullanın. Otofaji oldukça dinamik olduğu için, CLP&#39;nin IP enjeksiyonları gerçekleştirerek otofaj...

Watch this Scientific Journal Video about In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells at JoVE.com

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

Aktif otofaji, Kas Kök Hücresi (MuSC) aktivasyonu için gerekli olan üretken kas rejenerasyonu ile ilişkilidir. Burada, LC3&#39;ün in situ tespiti için bir protokol sunuyoruz, kontrol ve yaralı farelerdeki kas dokusu bölümlerinin MyoD pozitif MuSC&#39;lerinde bir otofaji markörü.

Otofajinin Kas Hücrelerinde Yerinde İmmunofloresan Boyanması

Increasing evidence points to autophagy as a crucial regulatory process to preserve tissue homeostasis. It is known that autophagy is involved in skeletal ...

The overall goal of this immunofluorescent staining protocol is to monitor the autophagic process in muscle satellite cells during skeletal muscle regeneration. This method can help answer key questions in the tissue regeneration field about understanding the contribution of autophagy in deriving muscle regeneration and monitoring autophagy in satellite cells. The main advantage of this technique is that the autophagic process can be directly detected in satellite cells in tissue section from unperturbed and damaged muscles.Demonstrating this procedure will be Francesco Castagnetti, a great student from my laboratory. To induce acute skeletal muscle injury, load an insulin syringe, equipped with a 30 gauge needle, with 20 microliters of sterile filtered 10 micromolar cardiotoxin stock. Inject the cardiotoxin directly into the left tibialis anterior muscle of an equal number of male and female two month old, approximately 20 gram, C57 black six mice.To assess the autophagic flux, starting at 24 hours post injury, administer 50 milligrams per kilogram chloroquine every 24 hours for four days by intraperitoneal injection. Chloroquine treatment is critical as it leads to LC3 accumulation and detectability under active autophagic flux conditions. Five days after the injury, wet the animals with 70%ethanol.And use scissors to make a three millimeter perpendicular incision in the dorsal skin of the mouse at the hip level. Pull the skin towards the tail to expose the underlying muscles. And identify the tibialis anterior muscle.Use forceps to grasp the tibialis anterior muscle by the tendon. And carefully pull the muscle up toward the knee. Then pull the two distal tendons in opposite directions to separate the tibialis anterior muscle from the extensor digitorum longus muscle and cut the proximal tendon.Use the forceps to remove the thin fascia covering the muscle, taking care not to damage the tissue. And use a paper towel to remove any excess moisture from the sample. Place the excised muscle into the center of a tissue mold, containing optimal cutting temperature compound.And use pre-chilled forceps to carefully lower the mold into liquid nitrogen chilled isopentane for 20 to 30 seconds. Then store the frozen sample at negative 80 degrees celsius. After at least one night at negative 80 degrees celsius, set the cryostat between negative 15 to negative 23 degrees celsius and use a drop of optimal cutting temperature compound to attach the muscle sample to the cryostat chuck with the bulk of the tissue oriented at the top of the chuck.Then obtain seven to eight micrometer thick sections, collecting three to four tissue slices per slide. After all of the section have been acquired, air dry the tissue samples for 10 to 15 minutes, followed by hematoxylin and eosin staining. Check the quality of the sections under an optical microscope at a 10X magnification.And fix the samples with 4%paraformaldehyde in an incubation chamber. After 10 minutes, remove the fixative with four five to seven minute washes in PBS. Then use a hydrophobic pen to draw a barrier around the tissue sections.And cover the section with 200 microliters of negative 20 degree celsius methanol for a five minute incubation at negative 20 degrees celsius. Methanol permeabilization is critical as it facilitates a good retention of the LC3 staining. Next, wash the section four times in PBS as just demonstrated.And block the tissue samples with an unconjugated affinity purified anti-mouse IGG FAB fragment for at least 60 minutes at room temperature. At the end of the incubation, incubate the specimens in 100 microliters of primary antibody mix at four degrees celsius overnight. The next morning, wash the slides with 1%bovine serum albumin in PBS four times for 10 minutes per wash.Then incubate the samples with the appropriate secondary antibody cocktail for 45 minutes at room temperature protected from light. At the end of the secondary antibody incubation, wash the slides four times in PBS for five to seven minutes per wash. Followed by labeling with the second primary antibody cocktail for one to two hours at room temperature protected from light.Wash the sections in 1%BSA and PBS four times. Followed by incubation in the appropriate second antibody for 45 minutes at room temperature protected from light. Then wash the samples in PBS and label the tissues with 200 microliters of DAPI per slide for five minutes at room temperature protected from light.After one last set of PBS washes, remove the excess PBS from the slides and add 10 microliters of glycerol in PBS onto the center of each slide. Cover each slide with a coverslip, taking care to avoid bubbles. Select the 63X objective on a four laser confocal microscope integrated with an image capture system and analytical software.Center the fields of active regeneration on the first slide to allow manual focus of the regenerative areas. Then locate the myoD positive muscle satellite cells positioned within the laminin labeled myofibers. And obtain images within at least seven fields from at least three mice per experimental group.H and E staining reveals that while unperturbed muscles displayed generally invariable myofiber sizes, injured muscles typically featured disrupted myofibers that differ in size and are abundant in inflammatory infiltrate. Further, the central nucleation of the myofibers, that is absent in unperturbed muscles, is a sign of new fiber formation in mice undergoing active regeneration. MyoD positive muscle satellite cells express LC3 and laminin staining allows the proper localization of muscle cells in the niche beneath the basal lamina.While uninjured mice do not demonstrate any muscle satellite cell activation as evidenced by their lack of myoD positive cells and LC3 signal. Once mastered, this technique can be completed in two days if it is performed properly. While attempting this procedure, it is important to remember to always keep the tissue sections hydrated.After its development, this technique paved the way for researching the field of tissue regeneration to explore the autophagic process during regular and paralogical skeletal muscle regeneration in mice and human biopsies. After watching this video, you should have a good understanding of how to identify LC3 in satellite cells during muscle regeneration.

Research

JoVE Journal

Biology

In Situ Immunofluorescent Staining of Autophagy in Muscle Stem Cells

Procedures for Rat in situ Skeletal Muscle Contractile Properties

Procedures for Rat in situ Skeletal Muscle Contractile Properties

Rat Prosedürleri<em&gt; In situ</em&gt; İskelet Kası Kontraktil Özellikleri

There are many circumstances where it is desirable to obtain the contractile response of skeletal muscle under physiological circumstances: normal ...

Biyoloji

Phenotypic Analysis and Isolation of Murine Hematopoietic Stem Cells and Lineage-committed Progenitors

Mürin Hematopoetik Kök Hücreler ve Lineage-kararlı progenitörlerin Fenotipik Analizi ve İzolasyon

The bone marrow is the principal site where HSCs and more mature blood cells lineage progenitors reside and differentiate in an adult organism. HSCs ...

Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells (iPSCs) Using Retroviral Vector with GFP

Reprogramming Human Somatic Cells into Induced Pluripotent Stem Cells iPSCs Using Retroviral Vector with GFP

GFP Retroviral Vector kullanarak İsteyerek Pluripotent Kök Hücrelerinin (iPSCs) içine İnsan somatik hücreleri yeniden programlama

Human embryonic stem cells (hESCs) are pluripotent and an invaluable cellular sources for in vitro disease modeling and regenerative medicine1. It has ...

Evaluation of Muscle Function of the Extensor Digitorum Longus Muscle Ex vivo and Tibialis Anterior Muscle In situ in Mice

Evaluation of Muscle Function of the Extensor Digitorum Longus Muscle Ex vivo and Tibialis Anterior Muscle In situ in Mice

Ekstansör digitorum longus kas ve kas Fonksiyonunun Değerlendirilmesi<em&gt; Ex vivo</em&gt; Ve tibialis anterior kas<em&gt; Yerinde</em&gt; Farelerde

Body  movements are mainly provided by mechanical function of skeletal muscle. Skeletal  muscle is composed of numerous bundles of myofibers that are ...

Isolation, Culture, and Transplantation of Muscle Satellite Cells

Kas Uydu Hücreleri İzolasyon, Kültür ve Transplantasyon

Muscle satellite cells are a stem cell population required for postnatal skeletal muscle development and regeneration, accounting for 2-5% of sublaminal ...

Efficient Generation Human Induced Pluripotent Stem Cells from Human Somatic Cells with Sendai-virus

Sendai-virüs ile İnsan Somatik Verimli Üretimi İnsan uyarılmış pluripotent kök hücrelerin

A few years ago, the establishment of human induced pluripotent stem cells (iPSCs) ushered in a new era in biomedicine. Potential uses of human iPSCs ...

Profiling Individual Human Embryonic Stem Cells by Quantitative RT-PCR

Kantitatif RT-PCR ile Bireysel İnsan Embriyonik Kök Hücreleri profilleme

Heterogeneity of stem cell population hampers detailed understanding of stem cell biology, such as their differentiation propensity toward different ...

Nephrotoxin Microinjection in Zebrafish to Model Acute Kidney Injury

Zebra balığı Nephrotoxin Mikroenjeksiyon Akut Böbrek Yaralanması Model

The kidneys are susceptible to harm from exposure to chemicals they filter from the bloodstream. This can lead to organ injury associated with a rapid ...

A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions

Dinamik Hücresel Protrüzyonlarda Protein Konsantrasyonunun Yazılım Destekli Takibi için Grafiksel Bir Kullanıcı Arayüzü

Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex ...

In Situ Monitoring of Transiently Formed Molecular Chaperone Assemblies in Bacteria, Yeast, and Human Cells

Bakteri, Maya ve İnsan Hücrelerinde Geçici Olarak Oluşan Moleküler Şaperon Meclislerinin Yerinde İzlenmesinde

J-domain proteins (JDPs) form the largest and the most diverse co-chaperone family in eukaryotic cells. Recent findings show that specific members of the ...