AP20187isacell-permeablemoleculeusedtodimerizeFK506-bindingprotein(FKBP)fusionproteinsandinitiatebiologicalsignalingcascadesandgeneexpressionordisruptprotein-proteininteractions.

FKBPhomodimerizer^[1]

WhenLNCaPcellsaretreatedwithAP20187(100nM),ro-iCaspase-9levelsaresignificantlyreduced,andthesmallerprocessedactivecaspase-9becomesapparent^[2].

Real-timePCRanalysisshowsthatAP20187(0.5mg/kg,2mg/kg,or5mg/kg)treatmentsignificantlyincreasesthelevelsofCHOPmRNAintheCNSofPLP/Fv2E-PERKmiceatPID12.AP20187treatmentsignificantlyalleviatesEAE-inducedmyelindamageinthesemice.AP20187treatmentsignificantlyreducesthenumberofdegeneratingaxonsandincreasesthedensityofaxonsinthedemyelinatinglesionsinthelumbarspinalcordofPLP/Fv2E-PERKmice^[2].

• [1].AhmedS,etal.PhotocleavabledimerizerfortherapidreversalofmoleculartrapantagoNISTs.JBiolChem.2014Feb21;289(8):4546-52.

  [2].LinW,etal.Oligodendrocyte-specificactivationofPERKsignalingprotectsmiceagainstexperimentalautoimmuneencephalomyelitis.JNeurosci.2013Apr3;33(14):5980-91.

  [3].HaasME,etal.TheRoleofProproteinConvertaseSubtilisin/KexinType9inNephroticSyndrome-AssociatedHypercholesterolemia.Circulation.2016Jul5;134(1):61-72.

AP20187isdissolvedinPBS^[2].

Fortheinvitrostudy,16hafterADVinfection,cellsaretreatedwithR1881(10nM),AP20187(10nM),both,orneitherfor8h.CellsarethenrinsedwithPBSandfixedwith4%paraformaldehydefor1hatroomtemperature.AfterrinsingwithPBS,cellsareincubatedinice-coldpermeABIlizationsolution(0.1%TritonX-100,0.1%sodiumcitrate)for2minat0°C.CellsarerinsedwithPBSandstainedwithTUNELreactionmixturefor60minat37°C.AfteranotherPBSwash,cellsareincubatedwithConverter-APfor30minat37°C.Cellsarerinsedandincubatedwithsubstrate5-bromo-4-chloro-3-indolylphosphate/nitrobluetetrazoliumfor30min.AfterafinalPBSrinse(repeatedtwice),cellsaremicrophotographed^[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

AP20187isdissolvedin100%ethanolataconcentrationof62.5mg/mLandstored(20°C).Thentreatmentsolutionsisfreshlypreparedfromadilutionof1.25mg/mLconsistingof4%ethanol,10%PEG-400,and2%Tween-20inwater^[2].

Mice^[2]
ToactivatethetransgeneFv2E-PERKinoligodendrocytes,PLP/Fv2E-PERKtransgenicmicearegivenintraperitonealinjectionsofAP20187dailyatadoseof0.5mg/kg,2mg/kg,or5mg/kg.LyophilizedAP20187isdissolvedin100%ethanolataconcentrationof62.5mg/mLstocksolutionandstoredat−20°C.Injectionsolutionsconsistof4%ethanol,10%PEG-400,and2%Tween-20inwater.Thetransgenicmicereceivingonlythevehicle(4%ethanol,10%PEG-400,2%Tween-20inwater)servedascontrols.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

• [1].AhmedS,etal.Photocleavabledimerizerfortherapidreversalofmoleculartrapantagonists.JBiolChem.2014Feb21;289(8):4546-52.

  [2].LinW,etal.Oligodendrocyte-specificactivationofPERKsignalingprotectsmiceagainstexperimentalautoimmuneencephalomyelitis.JNeurosci.2013Apr3;33(14):5980-91.

  [3].HaasME,etal.TheRoleofProproteinConvertaseSubtilisin/KexinType9inNephroticSyndrome-AssociatedHypercholesterolemia.Circulation.2016Jul5;134(1):61-72.

1482.75

C₈₂H₁₀₇N₅O₂₀

195514-80-8

RoomtemperatureincontinentalUS;mayvaryelsewhere

DMSO:≥57mg/mL

AP20187isdissolvedin4%ethanol,10%PEG-400,1.75%Tween-20inH₂O^[3].

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

• [1].AhmedS,etal.Photocleavabledimerizerfortherapidreversalofmoleculartrapantagonists.JBiolChem.2014Feb21;289(8):4546-52.

  [2].LinW,etal.Oligodendrocyte-specificactivationofPERKsignalingprotectsmiceagainstexperimentalautoimmuneencephalomyelitis.JNeurosci.2013Apr3;33(14):5980-91.

  [3].HaasME,etal.TheRoleofProproteinConvertaseSubtilisin/KexinType9inNephroticSyndrome-AssociatedHypercholesterolemia.Circulation.2016Jul5;134(1):61-72.