Name: HIV-1感染T細胞における小胞体ストレスとアンフォールドタンパク質応答の測定とHIV-1複製におけるその役割の解析 (動画) | JoVE Journal
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Description: 学ぶHIV-1感染T細胞における小胞体ストレスとアンフォールドタンパク質応答の測定法。最適なUPR活性化閾値がウイルス複製に必要であることを明らかにし、包括的なプロトコルを提供する。

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<meta charset="utf8"></head><body>T細胞におけるHIV-1感染時の小胞体ストレスとUPR活性化の研究

<ol><li>Levy, J. A. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aLevy%2C+JA+%22HIV+and+the+Pathogenesis+of+AIDS.+3rd%22">HIV and the Pathogenesis of AIDS. 3rd</a>. , ASM Press. (2007).</li>
<li>Hebert, D. N., Molinari, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((In+and+out+of+the+ER%3A+Protein+folding%2C+quality+control%2C+degradation%2C+and+related+human+diseases%5BTitle%5D)+AND+%22Physiol+Rev%22%5BJournal%5D)">In and out of the ER: Protein folding, quality control, degradation, and related human diseases.</a> Physiol Rev. 87 (4), 1377-1408 (2007).</li>
<li>Mori, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+unfolded+protein+response%3A+The+dawn+of+a+new+field%5BTitle%5D)+AND+%22Proc+Jpn+Acad+Ser+B+Phys+Biol+Sci%22%5BJournal%5D)">The unfolded protein response: The dawn of a new field.</a> Proc Jpn Acad Ser B Phys Biol Sci. 91 (9), 469-480 (2015).</li>
<li>Balch, W. E., Morimoto, R. I., Dillin, A., Kelly, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Adapting+proteostasis+for+disease+intervention%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">Adapting proteostasis for disease intervention.</a> Science. 319 (5865), 916-919 (2008).</li>
<li>Kaufman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Orchestrating+the+unfolded+protein+response+in+health+and+disease%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">Orchestrating the unfolded protein response in health and disease.</a> J Clin Invest. 110 (10), 1389-1398 (2002).</li>
<li>Ron, D., Walter, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Signal+integration+in+the+endoplasmic+reticulum+unfolded+protein+response%5BTitle%5D)+AND+%22Nat+Rev+Mol+Cell+Biol%22%5BJournal%5D)">Signal integration in the endoplasmic reticulum unfolded protein response.</a> Nat Rev Mol Cell Biol. 8 (7), 519-529 (2007).</li>
<li>Marciniak, S. J., Garcia-Bonilla, L., Hu, J., Harding, H. P., Ron, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Activation-dependent+substrate+recruitment+by+the+eukaryotic+translation+initiation+factor+2+kinase+PERK%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Activation-dependent substrate recruitment by the eukaryotic translation initiation factor 2 kinase PERK.</a> J Cell Biol. 172 (2), 201-209 (2006).</li>
<li>Harding, H. P., Zhang, Y., Ron, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protein+translation+and+folding+are+coupled+by+an+endoplasmic-reticulum-resident+kinase%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase.</a> Nature. 397 (6716), 271-274 (1999).</li>
<li>Vattem, K. M., Wek, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reinitiation+involving+upstream+ORFs+regulates+ATF4+mRNA+translation+in+mammalian+cells%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Reinitiation involving upstream ORFs regulates ATF4 mRNA translation in mammalian cells.</a> Proc Natl Acad Sci U S A. 101 (31), 11269-11274 (2004).</li>
<li>Harding, H. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+integrated+stress+response+regulates+amino+acid+metabolism+and+resistance+to+oxidative+stress%5BTitle%5D)+AND+%22Mol+Cell%22%5BJournal%5D)">An integrated stress response regulates amino acid metabolism and resistance to oxidative stress.</a> Mol Cell. 11 (3), 619-633 (2003).</li>
<li>Novoa, I., Zeng, H., Harding, H. P., Ron, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Feedback+inhibition+of+the+unfolded+protein+response+by+GADD34-mediated+dephosphorylation+of+eIF2%CE%B1%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Feedback inhibition of the unfolded protein response by GADD34-mediated dephosphorylation of eIF2α.</a> J Cell Biol. 153 (5), 1011-1021 (2001).</li>
<li>Haze, K., Yoshida, H., Yanagi, H., Yura, T., Mori, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mammalian+transcription+factor+ATF6+is+synthesized+as+a+transmembrane+protein+and+activated+by+proteolysis+in+response+to+endoplasmic+reticulum+stress%5BTitle%5D)+AND+%22Mol+Biol+Cell%22%5BJournal%5D)">Mammalian transcription factor ATF6 is synthesized as a transmembrane protein and activated by proteolysis in response to endoplasmic reticulum stress.</a> Mol Biol Cell. 10 (11), 3787-3799 (1999).</li>
<li>Ye, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ER+stress+induces+cleavage+of+membrane-bound+ATF6+by+the+same+proteases+that+process+SREBPs%5BTitle%5D)+AND+%22Mol+Cell%22%5BJournal%5D)">ER stress induces cleavage of membrane-bound ATF6 by the same proteases that process SREBPs.</a> Mol Cell. 6 (6), 1355-1364 (2000).</li>
<li>Tirasophon, W., Welihinda, A. A., Kaufman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+stress+response+pathway+from+the+endoplasmic+reticulum+to+the+nucleus+requires+a+novel+bifunctional+protein+kinase%2Fendoribonuclease+%28Ire1p%29+in+mammalian+cells%5BTitle%5D)+AND+%22Genes+Dev%22%5BJournal%5D)">A stress response pathway from the endoplasmic reticulum to the nucleus requires a novel bifunctional protein kinase/endoribonuclease (Ire1p) in mammalian cells.</a> Genes Dev. 12 (12), 1812-1824 (1998).</li>
<li>Yoshida, H., Matsui, T., Yamamoto, A., Okada, T., Mori, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((XBP1+mRNA+is+induced+by+ATF6+and+spliced+by+IRE1+in+response+to+ER+stress+to+produce+a+highly+active+transcription+factor%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">XBP1 mRNA is induced by ATF6 and spliced by IRE1 in response to ER stress to produce a highly active transcription factor.</a> Cell. 107 (7), 881-891 (2001).</li>
<li>Calfon, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((IRE1+couples+endoplasmic+reticulum+load+to+secretory+capacity+by+processing+the+XBP-1+mRNA%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">IRE1 couples endoplasmic reticulum load to secretory capacity by processing the XBP-1 mRNA.</a> Nature. 415 (6867), 92-96 (2002).</li>
<li>Wu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ATF6%CE%B1+optimizes+long-term+endoplasmic+reticulum+function+to+protect+cells+from+chronic+stress%5BTitle%5D)+AND+%22Dev+Cell%22%5BJournal%5D)">ATF6α optimizes long-term endoplasmic reticulum function to protect cells from chronic stress.</a> Dev Cell. 13 (3), 351-364 (2007).</li>
<li>Shoulders, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Stress-independent+activation+of+XBP1s+and%2For+ATF6+reveals+three+functionally+diverse+ER+proteostasis+environments%5BTitle%5D)+AND+%22Cell+Rep%22%5BJournal%5D)">Stress-independent activation of XBP1s and/or ATF6 reveals three functionally diverse ER proteostasis environments.</a> Cell Rep. 3 (4), 1279-1292 (2013).</li>
<li>Oslowski, C. M., Urano, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Measuring+ER+stress+and+the+unfolded+protein+response+using+mammalian+tissue+culture+system%5BTitle%5D)+AND+%22Methods+Enzymol%22%5BJournal%5D)">Measuring ER stress and the unfolded protein response using mammalian tissue culture system.</a> Methods Enzymol. 490, 71-92 (2011).</li>
<li>Gupta, S., Samali, A., Fitzgerald, U., Deegan, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Methods+for+monitoring+endoplasmic+reticulum+stress+and+the+unfolded+protein+response%5BTitle%5D)+AND+%22Int+J+Cell+Biol%22%5BJournal%5D)">Methods for monitoring endoplasmic reticulum stress and the unfolded protein response.</a> Int J Cell Biol. 2010, 830307(2010).</li>
<li>Wang, M., Kaufman, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protein+misfolding+in+the+endoplasmic+reticulum+as+a+conduit+to+human+disease%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Protein misfolding in the endoplasmic reticulum as a conduit to human disease.</a> Nature. 529 (7586), 326-335 (2016).</li>
<li>Beriault, D. R., Werstuck, G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Detection+and+quantification+of+endoplasmic+reticulum+stress+in+living+cells+using+the+fluorescent+compound%2C+Thioflavin+T%5BTitle%5D)+AND+%22Biochim+Biophys+Acta%22%5BJournal%5D)">Detection and quantification of endoplasmic reticulum stress in living cells using the fluorescent compound, Thioflavin T.</a> Biochim Biophys Acta. 1833 (10), 2293-2301 (2013).</li>
<li>Tripathi, A., Iyer, K., Mitra, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((HIV-1+replication+requires+optimal+activation+of+the+unfolded+protein+response%5BTitle%5D)+AND+%22FEBS+Lett%22%5BJournal%5D)">HIV-1 replication requires optimal activation of the unfolded protein response.</a> FEBS Lett. 597 (23), 2908-2930 (2023).</li>
<li>Platt, E. J., Wehrly, K., Kuhmann, S. E., Chesebro, B., Kabat, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Effects+of+CCR5+and+CD4+cell+surface+concentrations+on+infections+by+macrophagetropic+isolates+of+human+immunodeficiency+virus+type+1%5BTitle%5D)+AND+%22J+Virol%22%5BJournal%5D)">Effects of CCR5 and CD4 cell surface concentrations on infections by macrophagetropic isolates of human immunodeficiency virus type 1.</a> J Virol. 72 (4), 2855(1998).</li>
<li>Gervaix, A., West, D., Leoni, L. M., Richman, D. D., Wong-Staal, F., Corbeil, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+new+reporter+cell+line+to+monitor+HIV+infection+and+drug+susceptibility+in+vitro%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">A new reporter cell line to monitor HIV infection and drug susceptibility in vitro.</a> Proc Natl Acad Sci U S A. 94 (9), 4653-4658 (1997).</li>
<li>Augustine, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cyclin+F%2FFBXO1+interacts+with+HIV-1+viral+infectivity+factor+%28Vif%29+and+restricts+progeny+virion+infectivity+by+ubiquitination+and+proteasomal+degradation+of+vif+protein+through+SCFcyclin+F+E3+ligase+machinery%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Cyclin F/FBXO1 interacts with HIV-1 viral infectivity factor (Vif) and restricts progeny virion infectivity by ubiquitination and proteasomal degradation of vif protein through SCFcyclin F E3 ligase machinery.</a> J Biol Chem. 292 (13), 5349-5363 (2017).</li>
<li>Shihan, M. H., Novo, S. G., Le Marchand, S. J., Wang, Y., Duncan, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+simple+method+for+quantitating+confocal+fluorescent+images%5BTitle%5D)+AND+%22Biochem+Biophys+Rep%22%5BJournal%5D)">A simple method for quantitating confocal fluorescent images.</a> Biochem Biophys Rep. 25, 100916(2021).</li>
<li>Treiman, M., Caspersen, C., Christensen, S. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+tool+coming+of+age%3A+Thapsigargin+as+an+inhibitor+of+sarco-endoplasmic+reticulum+Ca2%2B-ATPases%5BTitle%5D)+AND+%22Trends+Pharmacol+Sci%22%5BJournal%5D)">A tool coming of age: Thapsigargin as an inhibitor of sarco-endoplasmic reticulum Ca2+-ATPases.</a> Trends Pharmacol Sci. 19 (4), 131-135 (1998).</li>
<li>Moffat, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+lentiviral+RNAi+library+for+human+and+mouse+genes+applied+to+an+arrayed+viral+high-content+screen%5BTitle%5D)+AND+%22Cell%22%5BJournal%5D)">A lentiviral RNAi library for human and mouse genes applied to an arrayed viral high-content screen.</a> Cell. 124 (6), 1283-1298 (2006).</li>
<li>Sodroski, J., Rosen, C., Goh, W. C., Haseltine, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+Transcriptional+activator+protein+encoded+by+the+x-lor+region+of+the+human+T-cell+leukemia+virus%5BTitle%5D)+AND+%22Science%22%5BJournal%5D)">A Transcriptional activator protein encoded by the x-lor region of the human T-cell leukemia virus.</a> Science. 228 (4706), 1430-1434 (1985).</li>
<li>Rosen, C. A., Sodroski, J. G., Kettman, R., Haseltine, W. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Activation+of+enhancer+sequences+in+type+II+human+T-cell+leukemia+virus+and+bovine+leukemia+virus+long+terminal+repeats+by+virus-associated+trans-acting+regulatory+factors%5BTitle%5D)+AND+%22J+Virol%22%5BJournal%5D)">Activation of enhancer sequences in type II human T-cell leukemia virus and bovine leukemia virus long terminal repeats by virus-associated trans-acting regulatory factors.</a> J Virol. 57 (3), 738-744 (1986).</li>
<li>Dandekar, D. H., Kumar, M., Ladha, J. S., Ganesh, K. N., Mitra, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+quantitative+method+for+normalization+of+transfection+efficiency+using+enhanced+green+fluorescent+protein%5BTitle%5D)+AND+%22Anal+Biochem%22%5BJournal%5D)">A quantitative method for normalization of transfection efficiency using enhanced green fluorescent protein.</a> Anal Biochem. 342 (2), 341-344 (2005).</li>
<li>Mahmood, T., Yang, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Western+blot%3A+Technique%2C+theory%2C+and+trouble+shooting%5BTitle%5D)+AND+%22N+Am+J+Med+Sci%22%5BJournal%5D)">Western blot: Technique, theory, and trouble shooting.</a> N Am J Med Sci. 4 (9), 429-434 (2012).</li>
<li>Ghosh, R., Gilda, J. E., Gomes, A. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+necessity+of+and+strategies+for+improving+confidence+in+the+accuracy+of+western+blots%5BTitle%5D)+AND+%22Expert+Rev+Proteomics%22%5BJournal%5D)">The necessity of and strategies for improving confidence in the accuracy of western blots.</a> Expert Rev Proteomics. 11 (5), 549-560 (2014).</li>
<li>Peña, J., Harris, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Dengue+virus+modulates+the+unfolded+protein+response+in+a+time-dependent+manner%5BTitle%5D)+AND+%22J+Biol+Chem%22%5BJournal%5D)">Dengue virus modulates the unfolded protein response in a time-dependent manner.</a> J Biol Chem. 286 (16), 14226-14236 (2011).</li>
<li>Soboleski, M. R., Oaks, J., Halford, W. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Green+fluorescent+protein+is+a+quantitative+reporter+of+gene+expression+in+individual+eukaryotic+cells%5BTitle%5D)+AND+%22FASEB+J%22%5BJournal%5D)">Green fluorescent protein is a quantitative reporter of gene expression in individual eukaryotic cells.</a> FASEB J. 19 (3), 440-442 (2005).</li>
<li>Martinez, Z. S., Castro, E., Seong, C. S., Cerón, M. R., Echegoyen, L., Llano, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fullerene+derivatives+strongly+inhibit+HIV-1+replication+by+affecting+virus+maturation+without+impairing+protease+activity%5BTitle%5D)+AND+%22Antimicrob+Agents+Chemother%22%5BJournal%5D)">Fullerene derivatives strongly inhibit HIV-1 replication by affecting virus maturation without impairing protease activity.</a> Antimicrob Agents Chemother. 60 (10), 5731-5741 (2016).</li>
<li>Askari, S., Javadpour, P., Rashidi, F. S., Dargahi, L., Kashfi, K., Ghasemi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Behavioral+and+molecular+effects+of+Thapsigargin-induced+brain+ER-+stress%3A+Encompassing+inflammation%2C+MAPK%2C+and+insulin+signaling+pathway%5BTitle%5D)+AND+%22Life%22%5BJournal%5D)">Behavioral and molecular effects of Thapsigargin-induced brain ER- stress: Encompassing inflammation, MAPK, and insulin signaling pathway.</a> Life. 12 (9), 1374(2022).</li>
<li>Jaskulska, A., Janecka, A. E., Gach-Janczak, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Thapsigargin-from+traditional+medicine+to+anticancer+drug%5BTitle%5D)+AND+%22Int+J+Mol+Sci%22%5BJournal%5D)">Thapsigargin-from traditional medicine to anticancer drug.</a> Int J Mol Sci. 22 (1), 4(2021).</li>
<li>Shaban, M. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Multi-level+inhibition+of+coronavirus+replication+by+chemical+ER+stress%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Multi-level inhibition of coronavirus replication by chemical ER stress.</a> Nat Commun. 12 (1), 5536(2021).</li>
<li>Marciniak, S. J., Chambers, J. E., Ron, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pharmacological+targeting+of+endoplasmic+reticulum+stress+in+disease%5BTitle%5D)+AND+%22Nat+Rev+Drug+Discov%22%5BJournal%5D)">Pharmacological targeting of endoplasmic reticulum stress in disease.</a> Nat Rev Drug Discov. 21 (2), 115-140 (2022).</li>
<li>Sriburi, R., Jackowski, S., Mori, K., Brewer, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((XBP1%3A+A+link+between+the+unfolded+protein+response%2C+lipid+biosynthesis%2C+and+biogenesis+of+the+endoplasmic+reticulum%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">XBP1: A link between the unfolded protein response, lipid biosynthesis, and biogenesis of the endoplasmic reticulum.</a> J Cell Biol. 167 (1), 35-41 (2004).</li>
<li>Siwecka, N., Rozpędek-Kamińska, W., Wawrzynkiewicz, A., Pytel, D., Diehl, J. A., Majsterek, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+structure%2C+activation+and+signaling+of+IRE1+and+its+role+in+determining+cell+fate%5BTitle%5D)+AND+%22Biomedicines%22%5BJournal%5D)">The structure, activation and signaling of IRE1 and its role in determining cell fate.</a> Biomedicines. 9 (2), 156(2021).</li>
</ol>

<table><tbody><tr><td>アクリルアミド</td><td>バイオラド、米国</td><td>1610107</td><td></td></tr><tr><td>Agarose</td><td>G-Biosciences、米国</td><td>RC1013</td><td></td></tr><tr><td>過硫酸アンモニウム</td><td>Sigma-Aldrich、米国</td><td>A3678</td><td></td></tr><tr><td>抗 ATF4 抗体</td><td>Cell Signaling Technology、米国</td><td>11815</td><td>ウェスタンブロット検出 希釈-1:1000</td></tr><tr><td>抗 ATF6 抗体</td><td>Abcam、英国</td><td>ab122897</td><td>ウェスタンブロット検出 Dilution-1:1000</td></tr><tr><td>anti-CHOP antibody</td><td>Cell Signaling Technology, USA</td><td>2897</td><td>ウェスタンブロット検出 Dilution-1:1000</td></tr><tr><td>anti-eIF2&alpha; antibody</td><td>Santa Cruz Biotechnology, USA</td><td>sc-11386</td><td>ウェスタンブロット検出 Dilution-1:2000&nbsp;</td></tr><tr><td>抗GADD34抗体</td><td>Abcam, UK</td><td>ab236516</td><td>ウェスタンブロット検出 希釈-1:1000</td></tr><tr><td>抗GAPDH抗体</td><td>Santa Cruz Biotechnology, USA</td><td>sc-32233</td><td>ウェスタンブロット検出 希釈-1:3000&nbsp;</td></tr><tr><td>抗HSPA5抗体</td><td>Cell Signaling Technology, USA</td><td>3177</td><td>Western blot detection Dilution-1:1000</td></tr><tr><td>anti-IRE1 antibody</td><td>Cell Signaling Technology, USA</td><td>3294</td><td>Western blot detection Dilution-1:2000</td></tr><tr><td>Anti-mouse HRP conjugate antibody&nbsp;</td><td>Biorad, USA</td><td>1706516</td><td>ウェスタンブロット検出 Dilution- 1:4000</td></tr><tr><td>anti-peIF2&alpha; antibody</td><td>Invitrogen, USA</td><td>44-728G</td><td>ウェスタンブロット検出 Dilution-1:1000</td></tr><tr><td>anti-PERK antibody</td><td>Cell Signaling Technology, USA</td><td>5683</td><td>ウェスタンブロット検出 Dilution-1:2000</td></tr><tr><td>抗 pIRE1 抗体</td><td>Abcam, UK</td><td>ab243665</td><td>ウェスタンブロット検出 Dilution-1:1000</td></tr><tr><td>抗pPERK抗体</td><td>Invitrogen, USA</td><td>PA5-40294</td><td>ウェスタンブロット検出 Dilution-1:2000</td></tr><tr><td>抗ウサギHRP標識抗体</td><td>Biorad, USA</td><td>1706515</td><td>ウェスタンブロット検出 Dilution- 1:4000</td></tr><tr><td>抗XBP1抗体</td><td>Abcam, UK</td><td>ab37152</td><td>ウェスタンブロット検出 Dilution-1:1000</td></tr><tr><td>ベンチトップ高速遠心分離機</td><td>Eppendorf, USA</td><td>5804R</td><td>ローター - F-45-30-11</td></tr><tr><td>ベンチトップ低速遠心分離機</td><td>エッペンドルフ(米国</td><td>) 5702R</td><td>ローター-A-4-38</td></tr><tr><td>ビスアクリルアミド</td><td>バイオラド(米国</td>) <td>1610201</td><td></td></tr><tr><td>ウシ血清アルブミン(BSA)</td><td>MPバイオメディカルズ(米国</td>) <td>160069</td><td></td></tr><tr><td>ブラッドフォード試薬</td><td>バイオラド(米国</td><td>) 5000006</td><td></td></tr><tr><td>CalPhos哺乳類トランスフェクションキット</td><td>Clontech(タカラバイオ)米国</td><td>631312</td><td>ウイルスストック調製</td></tr><tr><td>CEM-GFP</td><td>NIH, AIDS Repository, USA</td><td>3655</td><td></td></tr><tr><td>Clarity ECL substrate</td><td>Biorad, USA</td><td></td><td>1705061 化学発光検出基板</td></tr><tr><td>Clarity max ECL substrate</td><td>Biorad, USA</td><td>1705062</td><td>化学発光検出基板</td></tr><tr><td>共焦点レーザー走査顕微鏡</td><td>Olympus, Japan</td><td>Model:FV3000</td><td></td></tr><tr><td>Cytospin centrifuge</td><td>Thermo Fisher Scientific, USA</td><td>ASHA78300003</td><td></td></tr><tr><td>DMEM</td><td>Invitrogen, USA</td><td>11995073</td><td></td></tr><tr><td>DMSO</td><td>Sigma-Aldrich, USA</td><td>D2650</td><td></td></tr><tr><td>dNTPs</td><td>Promega, USA</td><td>U1515</td><td></td></tr><tr><td>DTT</td><td>Invitrogen, USA</td><td>R0861</td><td></td></tr><tr><td>EDTA</td><td>Invitrogen, USA</td><td>12635</td><td></td></tr><tr><td>EtBr</td><td>Invitrogen,</td> <td>USA'15585011</td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>Invitrogen, USA</td><td>16000044</td><td></td></tr><tr><td>G418</td><td>Invitrogen, USA</td><td>11811023</td><td></td></tr><tr><td>Glutaraldehyde 25%</td><td>Sigma-Aldrich, USA</td><td>G6257</td><td>Infectivity assay</td></tr><tr><td>Glycine</td><td>Thermo Fisher Scientific, USA</td><td>Q24755</td><td></td></tr><tr><td>HEK-293T</td><td>NCCS, India</td><td></td><td></td></tr><tr><td>HIV-1 Infectious Molecular Clone pNL4-3</td><td>NIH, AIDS Repository, USA</td><td>114</td><td></td></tr><tr><td>倒立顕微鏡</td><td>ニコン, 日本</td><td>モデル: Eclipse Ti2</td><td></td></tr><tr><td>iTaq Universal SYBR Green Supermix</td><td>Biorad, USA</td><td>1715124</td><td></td></tr><tr><td>Jurkat J6</td><td>NCCS, India</td><td></td><td></td></tr><tr><td>塩化マグネシウム</td><td>Sigma-Aldrich, USA</td><td>M8266</td><td>感染性アッセイ</td></tr><tr><td>MMLV-RT&nbsp;</td><td>Invitrogen, USA</td><td>28025013</td><td></td></tr><tr><td>MTT reagent</td><td>Sigma-Aldrich, USA</td><td>M5655</td><td>Cell viability assay</td></tr><tr><td>N,N-dimethyl formamide</td><td>Fluka Chemika</td><td>40255</td><td>Infectivity assay</td></tr><tr><td>NaCl</td><td>Thermo Fisher Scientific, USA</td><td>Q27605</td><td></td></tr><tr><td>NaF</td><td>Sigma-Aldrich, USA</td><td>201154</td><td></td></tr><tr><td>NP40</td><td>Invitrogen, USA</td><td>85124</td><td></td></tr><tr><td>P24抗原捕捉 ELISAキット</td><td>ABL, USA</td><td>5421</td><td></td></tr><tr><td>PageRuler prestained protein ladder</td><td>Sci-fi Biologicals, India</td><td>PGPMT078</td><td></td></tr><tr><td>Paraformaldehyde</td><td>Sigma-Aldrich, USA</td><td>P6148</td><td></td></tr><tr><td>pEGFP-N1</td><td>Clontech, USA</td><td>632515</td><td></td></tr><tr><td>Penicillin/Streptomycin</td><td>Invitrogen, USA</td><td>151140122</td><td></td></tr><tr><td>ホスファターゼ阻害剤</td><td>Sigma-Aldrich, USA</td><td>4906837001</td><td></td></tr><tr><td>Phusion High-fidelity PCR mastermix with GC buffer</td><td>NEB,USA</td><td>M05532</td><td></td></tr><tr><td>pLKO.1-TRC</td><td>Addgene, USA</td><td>10878</td><td>レンチウイルスクローニングベクター</td></tr><tr><td>pMD2.G</td><td>Addgene, USA</td><td>12259</td><td>VSV-G envelope</td></tr><tr><td>vector PMSF</td><td>Sigma-Aldrich, USA</td><td>P7626</td><td></td></tr><tr><td>ポリエチレンイミン(PEI)&nbsp;</td><td>Polysciences, Inc., USA</td><td>23966</td><td></td></tr><tr><td>フェリシアン化カリウム</td><td>Sigma-Aldrich, USA</td><td>244023</td><td>感染性アッセイ</td></tr><tr><td>フェロシアン化カリウム</td><td>Sigma-Aldrich, USA</td><td>P3289</td><td>感染性アッセイ</td></tr><tr><td>プロテアーゼ阻害剤</td><td>Sigma-Aldrich, USA&nbsp;</td><td></td><td></td>5056489001</tr><tr><td>psPAX2</td><td>Addgene, USA</td><td>12260</td><td>レンチウイルス包装プラスミド</td></tr><tr><td>Puromycin</td><td>Sigma-Aldrich, USA</td><td>P8833</td><td>安定細胞の選択</td></tr><tr><td>PVDF membrane</td><td>Biorad, USA</td><td>1620177</td><td></td></tr><tr><td>Random primers</td><td>Invitrogen, USA</td><td>48190011</td><td></td></tr><tr><td>RPMI 1640</td><td>Invitrogen, USA</td><td>22400105</td><td></td></tr><tr><td>SDS</td><td>Sigma-Aldrich, USA</td><td>L3771</td><td></td></tr><tr><td>Steady-Glo substrate</td><td>Promega, USA</td><td>E2510</td><td>Luciferase assay</td></tr><tr><td>T4 DNA ligase</td><td>Invitrogen, USA</td><td>15224017</td><td></td></tr><tr><td>TEMED</td><td>Invitrogen, USA</td><td>17919</td><td></td></tr><tr><td>Thapsigargin</td><td>Sigma-Aldrich, USA</td><td>T9033</td><td></td></tr><tr><td>Thioflavin T</td><td>Sigma-Aldrich, USA</td><td>596200</td><td></td></tr><tr><td>Tris</td><td>Thermo Fisher Scientific, USA</td><td>Q15965</td><td></td></tr><tr><td>Triton-X-100</td><td>Sigma-Aldrich, USA</td><td>T8787</td><td></td></tr><tr><td>Trizol</td><td>Invitrogen, USA</td><td>15596018</td><td></td></tr><tr><td>Tween 20</td><td>Sigma-Aldrich, USA</td><td>P1379</td><td></td></tr><tr><td>TZM-bl</td><td>NIH, AIDS Repository, USA</td><td>8129</td><td></td></tr><tr><td>超遠心分離機</td><td>BeckmanOptima L90K, USA</td><td>330049</td><td>Rotor-SW28Ti</td></tr><tr><td>UltraPure X-gal</td><td>Invitrogen, USA</td><td>15520-018</td><td>感染性アッセイ</td></tr></tbody></table>

hiv-1感染t細胞における小胞体ストレスとアンフォールドタンパク質応答の測定とhiv-1複製におけるその役割の解析

Measuring Endoplasmic Reticulum Stress and Unfolded Protein Response in HIV-1 Infected T-Cells and Analyzing its Role in HIV-1 Replication

HIV-1感染T細胞における小胞体ストレスとアンフォールドタンパク質応答の測定とHIV-1複製におけるその役割の解析

Viral infections can cause Endoplasmic Reticulum (ER) stress due to abnormal protein accumulation, leading to Unfolded Protein Response (UPR). Viruses ...

measuring-endoplasmic-reticulum-stress-unfolded-protein-response-hiv

Research

JoVE Journal

Immunology and Infection

免疫学と感染症

我々はTAT -応答LTR、HIV - 1 TAT -とRev依存的にレポーター遺伝子の発現を調節することができるREV...

緑色蛍光タンパク質を発現するリビジョンに依存するレンチウイルスベクターを用いたHIV - 1陽性細胞の特定のマーキング

この資料では、全内部反射蛍光（全反射）顕微鏡による平面脂質二重層をサポートしてガラス基板上にHIV-1エンベロープによって誘導されるウイルス学的シナプスの形成を可視化する方法について説明します。方法はまた、HIV-1エンベロープ誘発性ウイルス学的シナプス形成時に発生するシグナル伝達分子の活性化および再配布を検出する免疫​​蛍光染色と組み合わせることができます。...

合成脂質二重膜上でHIV-1エンベロープ誘発性ウイルス学的シナプスとシグナリングのイメージング

我々は、HIV-特異的CD4 T細胞によるサイトカイン分泌の調節における免疫調節経路の役割を調査するためのインビトロアッセイを開発した。...

インビトロアッセイで HIV特異的CD4 T細胞エフェクター機能に対する免疫調節経路の影響を評価する

無細胞HIV-1粒子とは異なり、感染したCD4 + T細胞を効果的に形質細胞様樹状細胞（pDC）によって感知されます。この原稿は、I型​​IFNの放出によって評価した末梢血単核細胞（PBMC）または単離されたpDCは、pDCにすることにより先天的センスを評価するためにHIV-1感染T細胞と共培養する方法が記載されています。...

HIV-1感染CD4の生得センシングを評価<sup&gt; +</sup&gt;使用して形質細胞様樹状細胞によるT細胞<em&gt;エクスビボ</em&gt;共培養系。

HIV-1複製サイクルの統合後のステップを標的とするRNA分子の有効性と毒性を評価する方法が説明されています。これらの方法は、新しい分子をスクリーニングし、既存の分子のフォーマットを最適化するのに役立ちます。...

遺伝子または薬物療法で使用するためのHIV-1産生を標的RNAの有効性および毒性の評価

この報告は、急性感染細胞の細胞質抽出物中のヒト免疫不全ウイルス1型(HIV-1)プレインテグレーションコンプレックス(PIC)関連統合活性を測定するためのin vitroアッセイについて説明しています。PIC関連HIV-1...

の測定インビトロ HIV-1プレインテグレーション複合体の統合活動

96 遺伝子の発現を量的に方法論が説明し、単一セルがex vivo、差動の同定を可能にする 18 の表面タンパク質は感染していない細胞を基準にしてウイルスに感染した細胞の遺伝子そして蛋白質を表明しました。+ T 研究 SIV 感染 CD4...

MRNA およびサル免疫不全ウイルス感染 cd4 表面タンパク質発現の単一細胞定量+ T 細胞孤立したアカゲザルから

初期 3' 末端の公平な測定だけでなく、単一座礁させた DNA の分子の変異プロファイルを提供するディープ シーケンス アプローチをご紹介します。メイン アプリケーションは、初期のレトロ ウイルス相補的 Dna (Cdna)、レトロ...

決定の 3' 末端と HIV-1 の間に初期の一本鎖のウイルス DNA 分子のシーケンスを逆に感染細胞における転写

ここでは、質量分析のためのHIV-1複製の存在下でのRev免疫沈殿について説明する。記載された方法は、HIV-1感染サイクルに関与するヌクレオラー因子の同定に使用することができ、研究中の経路の特徴付けのための他の疾患モデルに適用可能である。...

REV免疫沈殿と質量分析によるHIV-1複製中のヌクレオール因子の同定

ここでは、単一の細胞が急性事象およびナノ流体装置上の生産的なHIV-1感染を監視するプロトコルを提示する。イメージングデータは、ウイルスと宿主の受容体相互作用とシグナル伝達経路のダイナミクスを定義します。これは、ナノ流体ハイスループット縦単細胞培養およびイメージングにおけるシグナリング動態と分子相互作用を研究する最初の方法です。...

多パラメータ光流体プラットフォームを用いたカルシウム流入とHIV-1感染の単細胞特性

学ぶHIV-1感染T細胞における小胞体ストレスとアンフォールドタンパク質応答の測定法。最適なUPR活性化閾値がウイルス複製に必要であることを明らかにし、包括的なプロトコルを提供する。

HIV-1感染T細胞における小胞体ストレスとアンフォールドタンパク質応答の測定とHIV-1複製におけるその役割の解析

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