2001
Akt Down-regulation of p38 Signaling Provides a Novel Mechanism of Vascular Endothelial Growth Factor-mediated Cytoprotection in Endothelial Cells*
Gratton J, Morales-Ruiz M, Kureishi Y, Fulton D, Walsh K, Sessa W. Akt Down-regulation of p38 Signaling Provides a Novel Mechanism of Vascular Endothelial Growth Factor-mediated Cytoprotection in Endothelial Cells*. Journal Of Biological Chemistry 2001, 276: 30359-30365. PMID: 11387313, DOI: 10.1074/jbc.m009698200.Peer-Reviewed Original ResearchMeSH KeywordsAdenoviridaeAnimalsApoptosisBlotting, WesternCattleCell DeathCell LineCell SurvivalCells, CulturedDose-Response Relationship, DrugDown-RegulationEndothelial Growth FactorsEndothelium, VascularEnzyme ActivationEnzyme InhibitorsFlow CytometryHumansImidazolesLymphokinesMitogen-Activated Protein KinasesP38 Mitogen-Activated Protein KinasesPhosphatidylinositol 3-KinasesPhosphorylationProtein BindingProtein Serine-Threonine KinasesProto-Oncogene ProteinsProto-Oncogene Proteins c-aktPyridinesSignal TransductionTime FactorsUmbilical VeinsVascular Endothelial Growth Factor AVascular Endothelial Growth FactorsConceptsMEKK3 phosphorylationP38 activationMEKK3 kinase activityMitogen-activated protein kinaseP38 mitogen-activated protein kinaseP38-dependent apoptosisP38 MAPK inhibitor SB203580Dominant-negative RacInhibition of PIActivation of MKK3/6Vascular endothelial growth factorMAPK inhibitor SB203580P38 MAPK pathwayP38 MAPK activationEndothelial cellsEndothelial cell survivalGrowth factorRac activationProtein kinaseActive AktPro-apoptotic effectsKinase activityInhibitor SB203580MAPK activationP38 signalingHeat Shock Protein 90 Mediates the Balance of Nitric Oxide and Superoxide Anion from Endothelial Nitric-oxide Synthase*
Pritchard K, Ackerman A, Gross E, Stepp D, Shi Y, Fontana J, Baker J, Sessa W. Heat Shock Protein 90 Mediates the Balance of Nitric Oxide and Superoxide Anion from Endothelial Nitric-oxide Synthase*. Journal Of Biological Chemistry 2001, 276: 17621-17624. PMID: 11278264, DOI: 10.1074/jbc.c100084200.Peer-Reviewed Original ResearchConceptsEndothelial nitric oxide synthaseBovine coronary endothelial cellsNitric oxide synthaseHeat shock protein 90Shock protein 90Nitric oxidePhospho-eNOS levelsCoronary endothelial cellsProtein 90ENOS activityAssociation of hsp90Calcium ionophoreEndothelial cellsNitrite productionVascular biologySuperoxide anionAssociationPretreatmentHsp90SynthaseSphingosine 1-Phosphate Activates Akt, Nitric Oxide Production, and Chemotaxis through a GiProtein/Phosphoinositide 3-Kinase Pathway in Endothelial Cells*
Morales-Ruiz M, Lee M, Zöllner S, Gratton J, Scotland R, Shiojima I, Walsh K, Hla T, Sessa W. Sphingosine 1-Phosphate Activates Akt, Nitric Oxide Production, and Chemotaxis through a GiProtein/Phosphoinositide 3-Kinase Pathway in Endothelial Cells*. Journal Of Biological Chemistry 2001, 276: 19672-19677. PMID: 11278592, DOI: 10.1074/jbc.m009993200.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsBlotting, NorthernBlotting, WesternCattleCell MovementChemotaxisCulture Media, Serum-FreeDose-Response Relationship, DrugEndothelial Growth FactorsEndothelium, VascularEnzyme ActivationGenes, DominantGTP-Binding Protein alpha Subunits, Gi-GoLungLymphokinesLysophospholipidsNeovascularization, PhysiologicNitric OxideNitric Oxide SynthaseNitric Oxide Synthase Type IIIPhosphatidylinositol 3-KinasesPhosphorylationProtein BindingProtein Serine-Threonine KinasesProto-Oncogene ProteinsProto-Oncogene Proteins c-aktReceptors, Cell SurfaceSignal TransductionSphingosineTime FactorsVascular Endothelial Growth Factor AVascular Endothelial Growth FactorsVirulence Factors, BordetellaConceptsEndothelial differentiation gene familySerine/threonine kinase AktHeterotrimeric G proteinsThreonine kinase AktEDG-1 receptorGene familyAkt substrateKinase AktEndothelial cell chemotaxisActivates AktENOS phosphorylationAkt activationG proteinsCell survivalEndothelial nitric oxide synthasePhosphorylationAktCell chemotaxisSppSignalingGrowth factorVascular endothelial growth factorChemotaxisEndothelial cellsSphingosine
2000
Direct Interaction between Endothelial Nitric-oxide Synthase and Dynamin-2 IMPLICATIONS FOR NITRIC-OXIDE SYNTHASE FUNCTION*
Cao S, Yao J, McCabe T, Yao Q, Katusic Z, Sessa W, Shah V. Direct Interaction between Endothelial Nitric-oxide Synthase and Dynamin-2 IMPLICATIONS FOR NITRIC-OXIDE SYNTHASE FUNCTION*. Journal Of Biological Chemistry 2000, 276: 14249-14256. PMID: 11120737, DOI: 10.1074/jbc.m006258200.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAortaBlotting, WesternCalcimycinCattleCell LineDose-Response Relationship, DrugDynamin IDynaminsEndothelium, VascularGlutathione TransferaseGolgi ApparatusGTP PhosphohydrolasesIonophoresKineticsMicroscopy, ConfocalMicroscopy, FluorescenceNitric Oxide SynthaseNitric Oxide Synthase Type IIIPrecipitin TestsProtein BindingProtein BiosynthesisRatsRecombinant Fusion ProteinsTransfectionConceptsDynamin 2Bovine aortic endothelial cellsRecombinant eNOS proteinDirect protein-protein interactionDouble-label confocal immunofluorescenceProtein-protein interactionsSpecific protein interactionsDirect interactionClone 9 cellsEndothelial nitric oxide synthaseMembrane compartmentsLarge GTPaseGlutathione S-transferaseProtein interactionsNovel functionDynaminECV-304 cellsSynthase functionIntracellular signalsMembrane distributionConfocal immunofluorescenceFluorescent proteinENOS regulationGreen fluorescentProteinReconstitution of an Endothelial Nitric-oxide Synthase (eNOS), hsp90, and Caveolin-1 Complex in Vitro EVIDENCE THAT hsp90 FACILITATES CALMODULIN STIMULATED DISPLACEMENT OF eNOS FROM CAVEOLIN-1*
Gratton J, Fontana J, O'Connor D, Garcı́a-Cardeña G, McCabe T, Sessa W. Reconstitution of an Endothelial Nitric-oxide Synthase (eNOS), hsp90, and Caveolin-1 Complex in Vitro EVIDENCE THAT hsp90 FACILITATES CALMODULIN STIMULATED DISPLACEMENT OF eNOS FROM CAVEOLIN-1*. Journal Of Biological Chemistry 2000, 275: 22268-22272. PMID: 10781589, DOI: 10.1074/jbc.m001644200.Peer-Reviewed Original ResearchConceptsEndothelial nitric oxide synthaseAssociation of eNOSNitric oxide synthaseLung microvascular endothelial cellsCaveolin-1Microvascular endothelial cellsENOS enzymatic activityAction of CaMBovine lung microvascular endothelial cellsENOS functionCalcium-activated calmodulinConcentration of CaMShock protein 90Addition of CaMEndothelial cellsVitro EvidenceCav-1Protein 90AssociationPresence of Hsp90Vascular Endothelial Growth Factor–Stimulated Actin Reorganization and Migration of Endothelial Cells Is Regulated via the Serine/Threonine Kinase Akt
Morales-Ruiz M, Fulton D, Sowa G, Languino L, Fujio Y, Walsh K, Sessa W. Vascular Endothelial Growth Factor–Stimulated Actin Reorganization and Migration of Endothelial Cells Is Regulated via the Serine/Threonine Kinase Akt. Circulation Research 2000, 86: 892-896. PMID: 10785512, DOI: 10.1161/01.res.86.8.892.Peer-Reviewed Original ResearchConceptsSignal transduction mechanismsCell migrationActin reorganizationActive AktMyr-AktSerine/threonine kinase AktCell signal transduction mechanismsTransduction mechanismsThreonine kinase AktVascular endothelial growth factorDominant-negative AktDistinct signal transduction mechanismsBovine lung microvascular endothelial cellsEndothelial cellsAbsence of VEGFActin cytoskeletonGrowth factorMicrovascular endothelial cellsKinase AktActin rearrangementStress fibersF-actinCell locomotionEndothelial growth factorEndothelial cell proliferationEnhanced Electron Flux and Reduced Calmodulin Dissociation May Explain “Calcium-independent” eNOS Activation by Phosphorylation*
McCabe T, Fulton D, Roman L, Sessa W. Enhanced Electron Flux and Reduced Calmodulin Dissociation May Explain “Calcium-independent” eNOS Activation by Phosphorylation*. Journal Of Biological Chemistry 2000, 275: 6123-6128. PMID: 10692402, DOI: 10.1074/jbc.275.9.6123.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsCalciumCalmodulinCattleDimerizationEgtazic AcidElectronsEnzyme ActivationKineticsMutationNADH DehydrogenaseNADPNitric OxideNitric Oxide SynthaseNitric Oxide Synthase Type IIIPhosphorylationProtein Serine-Threonine KinasesProto-Oncogene ProteinsProto-Oncogene Proteins c-aktStatic ElectricityConceptsSerine 1179Reductase domainCalmodulin dissociationProtein kinase AktWild-type eNOSBovine endothelial nitric oxide synthaseEndothelial nitric oxide synthaseKinase AktRate-limiting stepReductase activityPhosphorylationENOS activationNOS functionPotential mechanismsAspartateENOS catalytic activityENOS activityCytochrome c reductionAktCalmodulinDomainProteinMutationsProductionActivityAngiopoietin-1 Inhibits Endothelial Cell Apoptosis via the Akt/Survivin Pathway*
Papapetropoulos A, Fulton D, Mahboubi K, Kalb R, O'Connor D, Li F, Altieri D, Sessa W. Angiopoietin-1 Inhibits Endothelial Cell Apoptosis via the Akt/Survivin Pathway*. Journal Of Biological Chemistry 2000, 275: 9102-9105. PMID: 10734041, DOI: 10.1074/jbc.275.13.9102.Peer-Reviewed Original ResearchMeSH KeywordsAngiopoietin-1AnimalsApoptosisCattleCells, CulturedEndothelium, VascularFlow CytometryInhibitor of Apoptosis ProteinsMembrane GlycoproteinsMicrotubule-Associated ProteinsNeoplasm ProteinsPhosphorylationProtein Serine-Threonine KinasesProteinsProto-Oncogene ProteinsProto-Oncogene Proteins c-aktSurvivinConceptsAkt/survivin pathwaySerine-threonine kinaseDeath-inducing stimuliPost-natal angiogenesisDominant-negative survivinEndothelial cellsEndothelial cell survivalAnti-apoptotic pathwaysSurvival machineryEndothelial cell apoptosisSurvivin pathwayApoptosis inhibitorCell survivalCell apoptosisVascular stabilizationAktSurvivinTie-2 receptorAngiogenic responseApoptosisCellsPathwayAngiogenesisMorphogenesisKinase
1999
Trafficking of Endothelial Nitric-oxide Synthase in Living Cells QUANTITATIVE EVIDENCE SUPPORTING THE ROLE OF PALMITOYLATION AS A KINETIC TRAPPING MECHANISM LIMITING MEMBRANE DIFFUSION*
Sowa G, Liu J, Papapetropoulos A, Rex-Haffner M, Hughes T, Sessa W. Trafficking of Endothelial Nitric-oxide Synthase in Living Cells QUANTITATIVE EVIDENCE SUPPORTING THE ROLE OF PALMITOYLATION AS A KINETIC TRAPPING MECHANISM LIMITING MEMBRANE DIFFUSION*. Journal Of Biological Chemistry 1999, 274: 22524-22531. PMID: 10428829, DOI: 10.1074/jbc.274.32.22524.Peer-Reviewed Original ResearchConceptsPlasma membraneENOS-GFPFluorescent protein fusion constructsProtein-protein interactionsRole of palmitoylationProtein fusion constructsLipid bilayersRate of traffickingEndothelial cell line ECV304Endothelial nitric oxide synthasePalmitoylation stateCellular domainsFusion constructsPerinuclear regionLiving cellsProtein diffusionFluorescence recoveryRegulation of eNOSMembrane markersPalmitoylationMutantsGolgiTraffickingMembrane diffusionEndothelial cellsRegulation of endothelium-derived nitric oxide production by the protein kinase Akt
Fulton D, Gratton J, McCabe T, Fontana J, Fujio Y, Walsh K, Franke T, Papapetropoulos A, Sessa W. Regulation of endothelium-derived nitric oxide production by the protein kinase Akt. Nature 1999, 399: 597-601. PMID: 10376602, PMCID: PMC3637917, DOI: 10.1038/21218.Peer-Reviewed Original ResearchConceptsProtein kinase AktKinase AktSerine/threonine protein kinase AktMutant eNOSRole of phosphorylationEndothelial nitric oxide synthaseSerine 1179Akt substrateSignal transductionGene transferAktAdenovirus-mediated gene transferPhosphorylationGrowth factorVascular endothelial growth factorEndothelial cellsRegulationSynthase isoformsEndothelial growth factorNitric oxide productionTransductionVascular remodellingOxide productionIsoformsProductionInduction of Nitric Oxide Synthase mRNA by Shear Stress Requires Intracellular Calcium and G-protein Signals and Is Modulated by PI 3 Kinase
Malek A, Jiang L, Lee I, Sessa W, Izumo S, Alper S. Induction of Nitric Oxide Synthase mRNA by Shear Stress Requires Intracellular Calcium and G-protein Signals and Is Modulated by PI 3 Kinase. Biochemical And Biophysical Research Communications 1999, 254: 231-242. PMID: 9920763, DOI: 10.1006/bbrc.1998.9921.Peer-Reviewed Original ResearchConceptsNitric oxide synthase mRNAPTX-sensitive G proteinsENOS mRNA levelsENOS mRNABovine aortic endothelial cellsIntracellular calciumPertussis toxinMRNA upregulationEndothelial nitric oxide synthase (eNOS) mRNAMRNA levelsEndothelin-1 mRNACalmodulin inhibitor WENOS gene promoterG proteinsSynthase mRNAAortic endothelial cellsTime-dependent increaseTyrosine kinase inhibitor herbimycin ACalcium entryBAPTA-AMInhibitor WEndothelial cellsTyrosine kinase activityMicrotubule integrityLaminar fluid shear stress
1998
Dynamic activation of endothelial nitric oxide synthase by Hsp90
García-Cardeña G, Fan R, Shah V, Sorrentino R, Cirino G, Papapetropoulos A, Sessa W. Dynamic activation of endothelial nitric oxide synthase by Hsp90. Nature 1998, 392: 821-824. PMID: 9580552, DOI: 10.1038/33934.Peer-Reviewed Original ResearchMeSH KeywordsAnimalsAntibiotics, AntineoplasticAortaBenzoquinonesCattleCell LineCOS CellsEndothelial Growth FactorsEndothelium, VascularEnzyme ActivationHistamineHSP90 Heat-Shock ProteinsHumansLactams, MacrocyclicLymphokinesMuscle RelaxationNitric OxideNitric Oxide SynthasePrecipitin TestsQuinonesRatsSignal TransductionStress, MechanicalTransfectionVascular Endothelial Growth Factor AVascular Endothelial Growth FactorsConceptsActivation of eNOSCellular targetsHeat shock protein 90Binding of HSP90Specific cellular targetsEndothelial nitric oxide synthaseMolecular chaperonesHsp90 associatesSignaling proteinsProtein foldingProtein 90Mechanotransduction pathwaysENOS complexG proteinsFluid shear stressHsp90Activation statePrecise roleGrowth factorDynamic activationVascular endothelial growth factorSynthaseNitric oxide synthaseEndothelial growth factorActivation
1997
Dissecting the Interaction between Nitric Oxide Synthase (NOS) and Caveolin FUNCTIONAL SIGNIFICANCE OF THE NOS CAVEOLIN BINDING DOMAININ VIVO *
Garcı́a-Cardeña G, Martasek P, Masters B, Skidd P, Couet J, Li S, Lisanti M, Sessa W. Dissecting the Interaction between Nitric Oxide Synthase (NOS) and Caveolin FUNCTIONAL SIGNIFICANCE OF THE NOS CAVEOLIN BINDING DOMAININ VIVO *. Journal Of Biological Chemistry 1997, 272: 25437-25440. PMID: 9325253, DOI: 10.1074/jbc.272.41.25437.Peer-Reviewed Original ResearchConceptsCaveolin-1Peripheral membrane proteinsInteraction of eNOSC-terminal tailAmino acids 310Direct interactionCo-transfection experimentsSite-directed mutagenesisNovel functional roleEndothelial nitric oxide synthaseMolecular chaperonesCytoplasmic domainCaveolin isoformsDeletion mutantsMammalian cellsEndothelial cell lysatesGlutathione S-transferaseMembrane proteinsCaveolin-2Coat proteinNegative regulationCaveolin-3Endothelial cellsDirect bindingGolgi regionCharacterization of bovine endothelial nitric oxide synthase as a homodimer with down-regulated uncoupled NADPH oxidase activity: tetrahydrobiopterin binding kinetics and role of haem in dimerization
LIST B, KLÖSCH B, VÖLKER C, GORREN A, SESSA W, WERNER E, KUKOVETZ W, SCHMIDT K, MAYER B. Characterization of bovine endothelial nitric oxide synthase as a homodimer with down-regulated uncoupled NADPH oxidase activity: tetrahydrobiopterin binding kinetics and role of haem in dimerization. Biochemical Journal 1997, 323: 159-165. PMID: 9173876, PMCID: PMC1218289, DOI: 10.1042/bj3230159.Peer-Reviewed Original ResearchSubstrate Binding and Calmodulin Binding to Endothelial Nitric Oxide Synthase Coregulate Its Enzymatic Activity
Presta A, Liu J, Sessa W, Stuehr D. Substrate Binding and Calmodulin Binding to Endothelial Nitric Oxide Synthase Coregulate Its Enzymatic Activity. Nitric Oxide 1997, 1: 74-87. PMID: 9701047, DOI: 10.1006/niox.1996.0110.Peer-Reviewed Original Research
1996
Elevated blood pressures in mice lacking endothelial nitric oxide synthase
Shesely E, Maeda N, Kim H, Desai K, Krege J, Laubach V, Sherman P, Sessa W, Smithies O. Elevated blood pressures in mice lacking endothelial nitric oxide synthase. Proceedings Of The National Academy Of Sciences Of The United States Of America 1996, 93: 13176-13181. PMID: 8917564, PMCID: PMC24066, DOI: 10.1073/pnas.93.23.13176.Peer-Reviewed Original ResearchMeSH KeywordsAnalysis of VarianceAnimalsBlood PressureCattleChimeraDNA PrimersEndothelium, VascularFemaleGenotypeHeterozygoteHypertensionIsoenzymesKidneyLipopolysaccharidesMaleMiceMice, Inbred C57BLMice, KnockoutNitric Oxide SynthasePolymerase Chain ReactionReninRNA, MessengerStem CellsTranscription, GeneticConceptsEndothelial nitric oxide synthaseBlood pressureNitric oxide synthaseHeart rateENOS proteinOxide synthaseENOS mutant miceLipopolysaccharide-induced deathElevated blood pressureNormal blood pressurePlasma renin concentrationBlood pressure regulationLower body weightKidney renin mRNAAnti-eNOS antibodiesAppropriate genetic controlsENOS locusENOS genotypesRenin concentrationVascular toneFemale miceRenin mRNAENOS geneENOS mutationBody weightEndothelial Nitric Oxide Synthase Is Regulated by Tyrosine Phosphorylation and Interacts with Caveolin-1*
García-Cardeña G, Fan R, Stern D, Liu J, Sessa W. Endothelial Nitric Oxide Synthase Is Regulated by Tyrosine Phosphorylation and Interacts with Caveolin-1*. Journal Of Biological Chemistry 1996, 271: 27237-27240. PMID: 8910295, DOI: 10.1074/jbc.271.44.27237.Peer-Reviewed Original ResearchConceptsNovel regulatory mechanismTyrosine phosphorylationCaveolin-1Bovine aortic endothelial cellsRegulatory mechanismsProtein tyrosine phosphatase inhibitorCaveolin-interacting proteinsPhosphoamino acid analysisTyrosine phosphatase inhibitorTreatment of BAECBovine lung microvascular endothelial cellsEndothelial nitric oxide synthaseSubcellular traffickingPhosphatase inhibitorCoat proteinEndothelial cellsMetabolic labelingSodium orthovanadatePhosphorylationCaveolaeAortic endothelial cellsLung microvascular endothelial cellsProteinAcid analysisImmunoprecipitationCharacterization of Bovine Endothelial Nitric Oxide Synthase Expressed inE. coli
Martasek P, Liu Q, Liu J, Roman L, Gross S, Sessa W, Masters B. Characterization of Bovine Endothelial Nitric Oxide Synthase Expressed inE. coli. Biochemical And Biophysical Research Communications 1996, 219: 359-365. PMID: 8604992, DOI: 10.1006/bbrc.1996.0238.Peer-Reviewed Original ResearchConceptsE. coliBaculovirus expression systemBovine endothelial nitric oxide synthaseSDS/PAGEHEK-293 cellsChaperonin GroELHeterologous expressionRecombinant enzymeExpression systemExpression vectorActive enzymeFormation assaysEnzymatic activityColiEnzymeTissue sourcesEndothelial constitutive nitric oxide synthaseProteinSingle bandSynthaseNitric oxide synthaseAbsorbance maximumEndothelial nitric oxide synthase
1995
The Golgi Association of Endothelial Nitric Oxide Synthase Is Necessary for the Efficient Synthesis of Nitric Oxide(∗)
Sessa W, Garca-Cardea G, Liu J, Keh A, Pollock J, Bradley J, Thiru S, Braverman I, Desai K. The Golgi Association of Endothelial Nitric Oxide Synthase Is Necessary for the Efficient Synthesis of Nitric Oxide(∗). Journal Of Biological Chemistry 1995, 270: 17641-17644. PMID: 7543089, DOI: 10.1074/jbc.270.30.17641.Peer-Reviewed Original ResearchConceptsHEK-293 cellsHeterologous expression systemEndothelial nitric oxide synthaseNovel GolgiGolgi associationExpression systemIntracellular signalsCultured endothelial cellsGolgiIntact blood vesselsEnzymeParticulate enzymeProteinEndothelial cellsCellsSynthaseNitric oxide synthaseOxide synthaseBlood vesselsCompartmentalizationNitric oxideTargetFunctional Analysis of the Human Endothelial Nitric Oxide Synthase Promoter SP1 AND GATA FACTORS ARE NECESSARY FOR BASAL TRANSCRIPTION IN ENDOTHELIAL CELLS (∗)
Zhang R, Min W, Sessa W. Functional Analysis of the Human Endothelial Nitric Oxide Synthase Promoter SP1 AND GATA FACTORS ARE NECESSARY FOR BASAL TRANSCRIPTION IN ENDOTHELIAL CELLS (∗). Journal Of Biological Chemistry 1995, 270: 15320-15326. PMID: 7541039, DOI: 10.1074/jbc.270.25.15320.Peer-Reviewed Original ResearchMeSH KeywordsAmino Acid OxidoreductasesAnimalsAortaBase SequenceBinding SitesCattleDNA PrimersDNA-Binding ProteinsEndothelium, VascularGATA2 Transcription FactorGene ExpressionHumansLuciferasesMolecular Sequence DataMutagenesis, Site-DirectedNitric Oxide SynthasePolymerase Chain ReactionPromoter Regions, GeneticRecombinant ProteinsRestriction MappingSp1 Transcription FactorTranscription FactorsTranscription, GeneticTransfectionConceptsBovine aortic endothelial cellsPromoter activitySp1 sitesBasal transcriptionGATA siteFurther deletionNuclear extractsENOS promoter activityAntibody supershift analysisDNA-protein complexesLuciferase activityENOS promoterLuc constructGel shift analysisSp-1 sitesBasal promoter activityLuciferase reporter geneSynthase gene expressionSp1 mutantGATA factorsNuclear proteinsGATA-2Mutant constructsReporter geneGel shift