CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells

doi:10.1007/s13238-018-0560-5

Protein Cell

2018, Vol. 9

Issue (11) : 945-965 https://doi.org/10.1007/s13238-018-0560-5

RESEARCH ARTICLE

CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells

Ping Wang^1,⁴, Zunpeng Liu^2,⁴, Xiaoqian Zhang^2,⁴, Jingyi Li^4,⁵, Liang Sun⁷, Zhenyu Ju⁸, Jian Li⁷, Piu Chan⁵, Guang-Hui Liu^1,^4,^5,^6,⁸(

), Weiqi Zhang^1,^4,⁵(

), Moshi Song^3,^4,⁶(

), Jing Qu^2,^4,⁶(

)

¹. National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China
². State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
³. State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China
⁴. University of Chinese Academy of Sciences, Beijing 100049, China
⁵. National Clinical Research Center for Geriatric Disorders, Xuanwu Hospital of Capital Medical University, Beijing 100053, China
⁶. Institute of Stem cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China
⁷. The MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of Gerontology, Beijing 100730, China
⁸. Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou 510632, China

Download: PDF(5589 KB)
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks

Abstract

Vascular cell functionality is critical to blood vessel homeostasis. Constitutive NF-κB activation in vascular cells results in chronic vascular inflammation, leading to various cardiovascular diseases. However, how NF-κB regulates human blood vessel homeostasis remains largely elusive. Here, using CRISPR/Cas9-mediated gene editing, we generated RelA knockout human embryonic stem cells (hESCs) and differentiated them into various vascular cell derivatives to study how NF-κB modulates human vascular cells under basal and inflammatory conditions. Multi-dimensional phenotypic assessments and transcriptomic analyses revealed that RelA deficiency affected vascular cells via modulating inflammation, survival, vasculogenesis, cell differentiation and extracellular matrix organization in a cell typespecific manner under basal condition, and that RelA protected vascular cells against apoptosis and modulated vascular inflammatory response upon tumor necrosis factor α (TNFα) stimulation. Lastly, further evaluation of gene expression patterns in IκBα knockout vascular cells demonstrated that IκBα acted largely independent of RelA signaling. Taken together, our data reveal a protective role of NF-κB/RelA in modulating human blood vessel homeostasis and map the human vascular transcriptomic landscapes for the discovery of novel therapeutic targets.

Keywords NF-κB RelA Stem cell Inflammation Apoptosis

Corresponding Author(s): Guang-Hui Liu,Weiqi Zhang,Moshi Song,Jing Qu

Issue Date: 29 November 2018

Cite this article:

Ping Wang,Zunpeng Liu,Xiaoqian Zhang, et al. CRISPR/Cas9-mediated gene knockout reveals a guardian role of NF-κB/RelA in maintaining the homeostasis of human vascular cells[J]. Protein Cell, 2018, 9(11): 945-965.

URL:

https://academic.hep.com.cn/pac/EN/10.1007/s13238-018-0560-5
https://academic.hep.com.cn/pac/EN/Y2018/V9/I11/945

1	Anders S, Pyl PT, Huber W (2015) HTSeq–a Python framework to work with high-throughput sequencing data. Bioinformatics 31:166–169 https://doi.org/10.1093/bioinformatics/btu638
2	Baker RG, Hayden MS, Ghosh S (2011) NF-kappaB, inflammation, and metabolic disease. Cell Metab 13:11–22 https://doi.org/10.1016/j.cmet.2010.12.008
3	Barnes PJ, Karin M (1997) Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases. N Engl J Med 336:1066–1071 https://doi.org/10.1056/NEJM199704103361506
4	Brand K, Page S, Rogler G, Bartsch A, Brandl R, Knuechel R, Page M, Kaltschmidt C, Baeuerle PA, Neumeier D (1996) Activated transcription factor nuclear factor-kappa B is present in the atherosclerotic lesion. J Clin Invest 97:1715–1722 https://doi.org/10.1172/JCI118598
5	Breitbach M, Kimura K, Luis TC, Fuegemann CJ, Woll PS, Hesse M, Facchini R, Rieck S, Jobin K, Reinhardt Jet al. (2018) In Vivo Labeling by CD73 Marks Multipotent Stromal Cells and Highlights Endothelial Heterogeneity in the Bone Marrow Niche. Cell Stem Cell 22(262–276):e267 https://doi.org/10.1016/j.stem.2018.01.008
6	Caplan AI, Correa D (2011) The MSC: an injury drugstore. Cell Stem Cell 9:11–15 https://doi.org/10.1016/j.stem.2011.06.008
7	Chen G, Chen Y, Chen H, Li L, Yao J, Jiang Q, Lin X, Wen J, Lin L (2011a) The effect of NF-kappaB pathway on proliferation and apoptosis of human umbilical vein endothelial cells induced by intermittent high glucose. Mol Cell Biochem 347:127–133 https://doi.org/10.1007/s11010-010-0620-5
8	Chen G, Qiao Y, Yao J, Jiang Q, Lin X, Chen F, Lin F, Lin M, Lin L, Zhu P (2011b) Construction of NF-kappaB-targeting RNAi adenovirus vector and the effect of NF-kappaB pathway on proliferation and apoptosis of vascular endothelial cells. Mol Biol Rep 38:3089–3094 https://doi.org/10.1007/s11033-010-9977-5
9	Chiba T, Kondo Y, Shinozaki S, Kaneko E, Ishigami A, Maruyama N, Umezawa K, Shimokado K (2006) A selective NFkappaB inhibitor, DHMEQ, reduced atherosclerosis in ApoE-deficient mice. J Atheroscler Thromb 13:308–313 https://doi.org/10.5551/jat.13.308
10	Courtois G, Smahi A, Reichenbach J, Doffinger R, Cancrini C, Bonnet M, Puel A, Chable-Bessia C, Yamaoka S, Feinberg Jet al. (2003) A hypermorphic IkappaBalpha mutation is associated with autosomal dominant anhidrotic ectodermal dysplasia and T cell immunodeficiency. J Clin Invest 112:1108–1115 https://doi.org/10.1172/JCI18714
11	da Silva Meirelles L, Caplan AI, Nardi NB (2008) In search of the in vivo identity of mesenchymal stem cells. Stem Cells 26:2287–2299 https://doi.org/10.1634/stemcells.2007-1122
12	Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA, Musunuru K (2013) Enhanced efficiency of human pluripotent stem cell genome editing through replacing TALENs with CRISPRs. Cell Stem Cell 12:393–394 https://doi.org/10.1016/j.stem.2013.03.006
13	Duan S, Yuan G, Liu X, Ren R, Li J, Zhang W, Wu J, Xu X, Fu L, Li Yet al. (2015) PTEN deficiency reprogrammes human neural stem cells towards a glioblastoma stem cell-like phenotype. Nat Commun 6:10068 https://doi.org/10.1038/ncomms10068
14	Fagerlund R, Behar M, Fortmann KT, Lin YE, Vargas JD, Hoffmann A (2015) Anatomy of a negative feedback loop: the case of IkappaBalpha. J R Soc Interface 12:0262 https://doi.org/10.1098/rsif.2015.0262
15	Fang J, Yang J, Wu X, Zhang G, Li T, Wang X, Zhang H, Wang CC, Liu GH, Wang L (2018) Metformin alleviates human cellular aging by upregulating the endoplasmic reticulum glutathione peroxidase. Aging Cell 7:e12765 https://doi.org/10.1111/acel.12765
16	Galley HF, Webster NR (2004) Physiology of the endothelium. Br J Anaesth 93:105–113 https://doi.org/10.1093/bja/aeh163
17	Gareus R, Kotsaki E, Xanthoulea S, van der Made I, Gijbels MJ, Kardakaris R, Polykratis A, Kollias G, de Winther MP, Pasparakis M (2008) Endothelial cell-specific NF-kappaB inhibition protects mice from atherosclerosis. Cell Metab 8:372–383 https://doi.org/10.1016/j.cmet.2008.08.016
18	Hajra L, Evans AI, Chen M, Hyduk SJ, Collins T, Cybulsky MI (2000) The NF-kappa B signal transduction pathway in aortic endothelial cells is primed for activation in regions predisposed to atherosclerotic lesion formation. Proc Natl Acad Sci USA 97:9052–9057 https://doi.org/10.1073/pnas.97.16.9052
19	Hiraoka A, Yano Ki K, Kagami N, Takeshige K, Mio H, Anazawa H, Sugimoto S (2001) Stem cell growth factor: in situ hybridization analysis on the gene expression, molecular characterization and in vitro proliferative activity of a recombinant preparation on primitive hematopoietic progenitor cells. Hematol J 2:307–315 https://doi.org/10.1038/sj.thj.6200118
20	Ijaz T, Wakamiya M, Sun H, Recinos A III, Tilton RG, Brasier AR (2016) Generation and characterization of a novel transgenic mouse harboring conditional nuclear factor-kappa B/RelA knockout alleles. BMC Dev Biol 16:32 https://doi.org/10.1186/s12861-016-0135-8
21	Jakkampudi A, Jangala R, Reddy BR, Mitnala S, Nageshwar Reddy D, Talukdar R (2016) NF-kappaB in acute pancreatitis: Mechanisms and therapeutic potential. Pancreatology 16:477–488 https://doi.org/10.1016/j.pan.2016.05.001
22	Janssen-Heininger YM, Poynter ME, Baeuerle PA (2000) Recent advances towards understanding redox mechanisms in the activation of nuclear factor kappaB. Free Radic Biol Med 28:1317–1327 https://doi.org/10.1016/S0891-5849(00)00218-5
23	Khan SY, Awad EM, Oszwald A, Mayr M, Yin X, Waltenberger B, Stuppner H, Lipovac M, Uhrin P, Breuss JM (2017) Premature senescence of endothelial cells upon chronic exposure to TNFα can be prevented by N-acetyl cysteine and plumericin. Sci Rep 7:39501 https://doi.org/10.1038/srep39501
24	Kida Y, Kobayashi M, Suzuki T, Takeshita A, Okamatsu Y, Hanazawa S, Yasui T, Hasegawa K (2005) Interleukin-1 stimulates cytokines, prostaglandin E2 and matrix metalloproteinase-1 production via activation of MAPK/AP-1 and NF-kappaB in human gingival fibroblasts. Cytokine 29:159–168 https://doi.org/10.1016/j.cyto.2004.10.009
25	Kim D, Langmead B, Salzberg SL (2015) HISAT: a fast spliced aligner with low memory requirements. Nat Methods 12:357–360 https://doi.org/10.1038/nmeth.3317
26	Kirton JP, Xu Q (2010) Endothelial precursors in vascular repair. Microvasc Res 79:193–199 https://doi.org/10.1016/j.mvr.2010.02.009
27	Kubben N, Zhang W, Wang L, Voss TC, Yang J, Qu J, Liu GH, Misteli T (2016) Repression of the antioxidant NRF2 pathway in premature aging. Cell 165:1361–1374 https://doi.org/10.1016/j.cell.2016.05.017
28	Kucharczak J, Simmons MJ, Fan Y, Gelinas C (2003) To be, or not to be: NF-kappaB is the answer–role of Rel/NF-kappaB in the regulation of apoptosis. Oncogene 22:8961–8982 https://doi.org/10.1038/sj.onc.1207230
29	Lee TH, Sottile J, Chiang HY (2015) Collagen inhibitory peptide R1R2 mediates vascular remodeling by decreasing inflammation and smooth muscle cell activation. PLoS ONE 10:e0117356 https://doi.org/10.1371/journal.pone.0117356
30	Li Y, Zhang W, Chang L, Han Y, Sun L, Gong X, Tang H, Liu Z, Deng H, Ye Yet al. (2016) Vitamin C alleviates aging defects in a stem cell model for Werner syndrome. Protein Cell 7:478–488 https://doi.org/10.1007/s13238-016-0278-1
31	Liu GH, Qu J, Shen X (2008) NF-kappaB/p65 antagonizes Nrf2-ARE pathway by depriving CBP from Nrf2 and facilitating recruitment of HDAC3 to MafK. Biochim Biophys Acta 1783:713–727 https://doi.org/10.1016/j.bbamcr.2008.01.002
32	Liu GH, Barkho BZ, Ruiz S, Diep D, Qu J, Yang SL, Panopoulos AD, Suzuki K, Kurian L, Walsh Cet al. (2011) Recapitulation of premature ageing with iPSCs from Hutchinson-Gilford progeria syndrome. Nature 472:221–225 https://doi.org/10.1038/nature09879
33	Liu GH, Qu J, Suzuki K, Nivet E, Li M, Montserrat N, Yi F, Xu X, Ruiz S, Zhang Wet al. (2012) Progressive degeneration of human neural stem cells caused by pathogenic LRRK2. Nature 491:603–607 https://doi.org/10.1038/nature11557
34	Love MI, Huber W, Anders S (2014) Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol 15:550 https://doi.org/10.1186/s13059-014-0550-8
35	Mallavia B, Recio C, Oguiza A, Ortiz-Munoz G, Lazaro I, Lopez-Parra V, Lopez-Franco O, Schindler S, Depping R, Egido Jet al. (2013) Peptide inhibitor of NF-kappaB translocation ameliorates experimental atherosclerosis. Am J Pathol 182:1910–1921 https://doi.org/10.1016/j.ajpath.2013.01.022
36	Morris O, Liu X, Domingues C, Runchel C, Chai A, Basith S, Tenev T, Chen H, Choi S, Pennetta Get al. (2016) Signal integration by the IkappaB protein pickle shapes drosophila innate host defense. Cell Host Microbe 20:283–295 https://doi.org/10.1016/j.chom.2016.08.003
37	Nedeljkovic ZS, Gokce N, Loscalzo J (2003) Mechanisms of oxidative stress and vascular dysfunction. Postgrad Med J 79:195–199 https://doi.org/10.1136/pmj.79.930.195
38	Osborn L, Kunkel S, Nabel GJ (1989) Tumor necrosis factor alpha and interleukin 1 stimulate the human immunodeficiency virus enhancer by activation of the nuclear factor kappa B. Proc Natl Acad Sci USA 86:2336–2340 https://doi.org/10.1073/pnas.86.7.2336
39	Pan H, Guan D, Liu X, Li J, Wang L, Wu J, Zhou J, Zhang W, Ren R, Zhang Wet al. (2016) SIRT6 safeguards human mesenchymal stem cells from oxidative stress by coactivating NRF2. Cell Res 26:190–205 https://doi.org/10.1038/cr.2016.4
40	Patsch C, Challet-Meylan L, Thoma EC, Urich E, Heckel T, O’Sullivan JF, Grainger SJ, Kapp FG, Sun L, Christensen Ket al. (2015) Generation of vascular endothelial and smooth muscle cells from human pluripotent stem cells. Nat Cell Biol 17:994–1003 https://doi.org/10.1038/ncb3205
41	Perkins ND (2007) Integrating cell-signalling pathways with NFkappaB and IKK function. Nat Rev Mol Cell Biol 8:49–62 https://doi.org/10.1038/nrm2083
42	Perkins ND, Gilmore TD (2006) Good cop, bad cop: the different faces of NF-kappaB. Cell Death Diff 13:759–772 https://doi.org/10.1038/sj.cdd.4401838
43	Rudijanto A (2007) The role of vascular smooth muscle cells on the pathogenesis of atherosclerosis. Acta Medica Indones 39:86–93
44	Salminen A, Huuskonen J, Ojala J, Kauppinen A, Kaarniranta K, Suuronen T (2008) Activation of innate immunity system during aging: NF-kB signaling is the molecular culprit of inflamm-aging. Ageing Res Rev 7:83–105 https://doi.org/10.1016/j.arr.2007.09.002
45	Schober A, Weber C (2005) Mechanisms of monocyte recruitment in vascular repair after injury. Antioxid Redox Signal 7:1249–1257 https://doi.org/10.1089/ars.2005.7.1249
46	Simeonidis S, Stauber D, Chen G, Hendrickson WA, Thanos D (1999) Mechanisms by which IkappaB proteins control NFkappaB activity. Proc Natl Acad Sci USA 96:49–54 https://doi.org/10.1073/pnas.96.1.49
47	Tas SW, Vervoordeldonk MJ, Tak PP (2009) Gene therapy targeting nuclear factor-kappaB: towards clinical application in inflammatory diseases and cancer. Curr Gene Therapy 9:160–170 https://doi.org/10.2174/156652309788488569
48	Tilstra JS, Clauson CL, Niedernhofer LJ, Robbins PD (2011) NFkappaB in Aging and Disease. Aging Dis 2:449–465
49	Uccelli A, Moretta L, Pistoia V (2008) Mesenchymal stem cells in health and disease. Nat Rev Immunol 8:726–736 https://doi.org/10.1038/nri2395
50	Wang L, Yi F, Fu L, Yang J, Wang S, Wang Z, Suzuki K, Sun L, Xu X, Yu Yet al. (2017) CRISPR/Cas9-mediated targeted gene correction in amyotrophic lateral sclerosis patient iPSCs. Protein Cell 8:365–378 https://doi.org/10.1007/s13238-017-0397-3
51	Wang S, Hu B, Ding Z, Dang Y, Wu J, Li D, Liu X, Xiao B, Zhang W, Ren Ret al. (2018) ATF6 safeguards organelle homeostasis and cellular aging in human mesenchymal stem cells. Cell Discov 4:2 https://doi.org/10.1038/s41421-017-0003-0
52	Wu Z, Zhang W, Song M, Wang W, Wei G, Li W, Lei J, Huang Y, Sang Y, Chan Pet al. (2018) Differential stem cell aging kinetics in Hutchinson-Gilford progeria syndrome and Werner syndrome. Protein Cell 9:333–350 https://doi.org/10.1007/s13238-018-0517-8
53	Yang J, Li J, Suzuki K, Liu X, Wu J, Zhang W, Ren R, Zhang W, Chan P, Izpisua Belmonte JCet al. (2017) Genetic enhancement in cultured human adult stem cells conferred by a single nucleotide recoding. Cell Res 27:1178–1181 https://doi.org/10.1038/cr.2017.86
54	Yu G, Wang LG, Han Y, He QY (2012) clusterProfiler: an R package for comparing biological themes among gene clusters. Omics 16:284–287 https://doi.org/10.1089/omi.2011.0118
55	Yue R, Shen B, Morrison SJ (2016) Clec11a/osteolectin is an osteogenic growth factor that promotes the maintenance of the adult skeleton. eLife 5 https://doi.org/10.7554/eLife.18782
56	Zhang W, Li J, Suzuki K, Qu J, Wang P, Zhou J, Liu X, Ren R, Xu X, Ocampo Aet al. (2015) Aging stem cells. A Werner syndrome stem cell model unveils heterochromatin alterations as a driver of human aging. Science 348:1160–1163 https://doi.org/10.1126/science.aaa1356
57	Zhang W, Song M, Qu J, Liu G (2018) Epigeneic modifications in cardiovascular aging and diseases. Circ Res (in press) https://doi.org/10.1161/CIRCRESAHA.118.312497

[1]

PAC-0945-18288-LGH_suppl_1

Download

[1]	Ermin Li, Xiuya Li, Jie Huang, Chen Xu, Qianqian Liang, Kehan Ren, Aobing Bai, Chao Lu, Ruizhe Qian, Ning Sun. BMAL1 regulates mitochondrial fission and mitophagy through mitochondrial protein BNIP3 and is critical in the development of dilated cardiomyopathy[J]. Protein Cell, 2020, 11(9): 661-679.
[2]	Kun Liu, Jiani Cao, Xingxing Shi, Liang Wang, Tongbiao Zhao. Cellular metabolism and homeostasis in pluripotency regulation[J]. Protein Cell, 2020, 11(9): 630-640.
[3]	Shijia Bi, Zunpeng Liu, Zeming Wu, Zehua Wang, Xiaoqian Liu, Si Wang, Jie Ren, Yan Yao, Weiqi Zhang, Moshi Song, Guang-Hui Liu, Jing Qu. SIRT7 antagonizes human stem cell aging as a heterochromatin stabilizer[J]. Protein Cell, 2020, 11(7): 483-504.
[4]	Matthieu Talagas, Nicolas Lebonvallet, François Berthod, Laurent Misery. Lifting the veil on the keratinocyte contribution to cutaneous nociception[J]. Protein Cell, 2020, 11(4): 239-250.
[5]	Rui Fu, Dawei Yu, Jilong Ren, Chongyang Li, Jing Wang, Guihai Feng, Xuepeng Wang, Haifeng Wan, Tianda Li, Libin Wang, Ying Zhang, Tang Hai, Wei Li, Qi Zhou. Domesticated cynomolgus monkey embryonic stem cells allow the generation of neonatal interspecies chimeric pigs[J]. Protein Cell, 2020, 11(2): 97-107.
[6]	Ruyi Xu, Yi Li, Yang Liu, Jianwei Qu, Wen Cao, Enfan Zhang, Jingsong He, Zhen Cai. How are MCPIP1 and cytokines mutually regulated in cancer-related immunity?[J]. Protein Cell, 2020, 11(12): 881-893.
[7]	Hua Qin, Andong Zhao. Mesenchymal stem cell therapy for acute respiratory distress syndrome: from basic to clinics[J]. Protein Cell, 2020, 11(10): 707-722.
[8]	Xuemei Fu, Shouhai Wu, Bo Li, Yang Xu, Jingfeng Liu. Functions of p53 in pluripotent stem cells[J]. Protein Cell, 2020, 11(1): 71-78.
[9]	Feng Li, Yuanlong Ge, Dan Liu, Zhou Songyang. The role of telomere-binding modulators in pluripotent stem cells[J]. Protein Cell, 2020, 11(1): 60-70.
[10]	Hui Cheng, Zhaofeng Zheng, Tao Cheng. New paradigms on hematopoietic stem cell differentiation[J]. Protein Cell, 2020, 11(1): 34-44.
[11]	Si Wang, Zheying Min, Qianzhao Ji, Lingling Geng, Yao Su, Zunpeng Liu, Huifang Hu, Lixia Wang, Weiqi Zhang, Keiichiro Suzuiki, Yu Huang, Puyao Zhang, Tie-Shan Tang, Jing Qu, Yang Yu, Guang-Hui Liu, Jie Qiao. Rescue of premature aging defects in Cockayne syndrome stem cells by CRISPR/Cas9-mediated gene correction[J]. Protein Cell, 2020, 11(1): 1-22.
[12]	Lili Yu, Kai-yuan Ji, Jian Zhang, Yanxia Xu, Yue Ying, Taoyi Mai, Shuxiang Xu, Qian-bing Zhang, Kai-tai Yao, Yang Xu. Core pluripotency factors promote glycolysis of human embryonic stem cells by activating GLUT1 enhancer[J]. Protein Cell, 2019, 10(9): 668-680.
[13]	Xing Zhang, Zunpeng Liu, Xiaoqian Liu, Si Wang, Yiyuan Zhang, Xiaojuan He, Shuhui Sun, Shuai Ma, Ng Shyh-Chang, Feng Liu, Qiang Wang, Xiaoqun Wang, Lin Liu, Weiqi Zhang, Moshi Song, Guang-Hui Liu, Jing Qu. Telomere-dependent and telomereindependent roles of RAP1 in regulating human stem cell homeostasis[J]. Protein Cell, 2019, 10(9): 649-667.
[14]	Lingling Geng, Zunpeng Liu, Weiqi Zhang, Wei Li, Zeming Wu, Wei Wang, Ruotong Ren, Yao Su, Peichang Wang, Liang Sun, Zhenyu Ju, Piu Chan, Moshi Song, Jing Qu, Guang-Hui Liu. Chemical screen identifies a geroprotective role of quercetin in premature aging[J]. Protein Cell, 2019, 10(6): 417-435.
[15]	Daisuke Aki, Qian Li, Hui Li, Yun-Cai Liu, Jee Ho Lee. Immune regulation by protein ubiquitination: roles of the E3 ligases VHL and Itch[J]. Protein Cell, 2019, 10(6): 395-404.

Viewed

Full text

Abstract

Cited

Shared

Discussed