Establishment of Inducible Wild Type and Mutant Myocilin-GFP-Expressing RGC5 Cell Lines 英文参考文献.docVIP

下载本文档

4
0
约5.55万字
约 12页
2017-05-11 发布于上海
举报
版权申诉

Establishment of Inducible Wild Type and Mutant Myocilin-GFP-Expressing RGC5 Cell Lines 英文参考文献.doc

1、本文档共12页，可阅读全部内容。
2、原创力文档（book118）网站文档一经付费（服务费），不意味着购买了该文档的版权，仅供个人/单位学习、研究之用，不得用于商业用途，未经授权，严禁复制、发行、汇编、翻译或者网络传播等，侵权必究。
3、本站所有内容均由合作方或网友上传，本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺！文档内容仅供研究参考，付费前请自行鉴别。如您付费，意味着您自己接受本站规则且自行承担风险，本站不退款、不进行额外附加服务；查看《如何避免下载的几个坑》。如果您已付费下载过本站文档，您可以点击这里二次下载。
4、如文档侵犯商业秘密、侵犯著作权、侵犯人身权等，请点击“版权申诉”（推荐），也可以打举报电话：400-050-0827(电话支持时间：9:00-18:30)。
5、该文档为VIP文档，如果想要下载，成为VIP会员后，下载免费。
6、成为VIP后，下载本文档将扣除1次下载权益。下载后，不支持退款、换文档。如有疑问请联系我们。
7、成为VIP后，您将拥有八大权益，权益包括：VIP文档下载权益、阅读免打扰、文档格式转换、高级专利检索、专属身份标志、高级客服、多端互通、版权登记。
8、VIP文档为合作方或网友上传，每下载1次，网站将根据用户上传文档的质量评分、类型等，对文档贡献者给予高额补贴、流量扶持。如果你也想贡献VIP文档。上传文档

Establishment of Inducible Wild Type and Mutant Myocilin-GFP-Expressing RGC5 Cell Lines 英文参考文献

EstablishmentofInducibleWildTypeandMutant Myocilin-GFP-ExpressingRGC5CellLines HongyuYing,XiangShen,BeatriceY.J.T.Yue* DepartmentofOphthalmologyandVisualSciences,UniversityofIllinoisatChicago,CollegeofMedicine,Chicago,Illinois,UnitedStatesofAmerica Abstract Background: Myocilin is a gene linked directly to juvenile- and adult-onset open angle glaucoma. Mutations including Gln368stop(Q368X)andPro370Leu(P370L)havebeenidentifiedinpatients.Theexactroleofmyocilinanditsfunctional associationwithglaucomaarestillunclear.Inthepresentstudy,weestablishedtetracycline-inducible(Tet-on)wildtypeand mutant myocilin-green fluorescence protein (GFP) expressing RGC5 stable cell lines and studied the changes in cell migrationandbarrierfunctionuponinduction. Methodology/PrincipalFindings:Afterseveralroundsofselection,clonesthatdisplayedlow,moderate,orhighexpression ofwildtype,Q368XorP370Lmyocilin-GFPupondoxycycline(Dox)inductionwereobtained.Thelevelsofwildtypeand mutantmyocilin-GFPinvariouscloneswereconfirmedbyWesternblotting.Comparedtonon-inducedcontrols,thecell migrationwasretarded,theactinstressfiberswerefewerandshorter,andthetrypsinizationtimeneededforcellstoround upwasreducedwhenwildtypeormutantmyocilinwasexpressed.Thebarrierfunctionwasinadditionaberrantfollowing induced expression of wild type, Q368X or P370L myocilin. Immunoblotting further showed that tight junction protein occludinwasdownregulatedininducedcells. Conclusions/Significance:Tet-oninducible,stableRGC5celllineswereestablished.Thesecelllines,expressingwildtypeor mutant(Q368XorP370L)myocilin-GFPuponDoxinduction,arevaluableinfacilitatingstudiessuchasproteomics,aswell as functional and pathogenesis investigations of disease-associated myocilin mutants. The barrier function was found impairedandthemigrationofcellswashinderedwithinducedexpressionofwildtypeandmutantmyocilininRGC5cell lines.Thereductioninbarrierfunctionmightberelatedtothedeclinedlevelofoccludin.Theretardedcellmigrationwas consistent with demonstrated myoci