Gear|Products|Items|Tools|Supplies|Devices|Machines|Accessories|Stuff|Goods|Gadgets|Outfits|Things|Materials|Objects|Appliances|Resources|Solutions|In

xperimentsusingdifferentiallytagged
proteins (Supplementary Fig. 3a,b). Domain mapping of DVL2
31
mutants
additional showed the C-terminal aspect of DVL2
such as the DEP domain was demanded for PTEN interaction
(Supplementary Fig. 3c,d). Endogenous PTEN and DVL2 also
interacted in a human retinal epithelial cell line (hTERT-RPE1)
that assembles cilia Gear|Products|Items|Tools|Supplies|Devices|Machines|Accessories|Stuff|Goods|Gadgets|Outfits|Things|Materials|Objects|Appliances|Resources|Solutions|Instruments|Components|Focus|Units} And Formation Throughout Vegas :Ruxolitinib Leaves With No See You Later in vitro (Fig. 3b). Also, endogenous
PTEN partially co-localized with ectopically expressed DVL in
areas adjacent to BBs in hTERT-RPE1 (data not shown) and
with Xenopus Dishevelled (Dsh) in MCCs (Supplementary
Fig. 2c). To much better visualize co-localization between PTEN and
DshinXenopusMCCs,wealsousedaDVLmutantlackingthe
DIXdomain(dDIX-Dsh).Thismutantretainsproperlocalization
totheBBareaanddoesnotformaggregates,whicharefrequently
20
detected when WT Dsh is overexpressed .

Interestingly, in
XenopusMCCsPTENanddDIX-Dsh-GFPwerelocalizednextto
BBs marked by Centrin-RFP and had been commonly observed in
adjacent regions that partially overlapped. Consequently, overlapping
PTENandDshsignals(SupplementaryFig.2c,whitearrows),as
very well as adjacent localization (magenta arrows) have been readily
obvious.
Previousreportsindicatethatciliadisassemblyistriggeredby
casein Gear|Products|Items|Tools|Supplies|Devices|Machines|Accessories|Stuff|Goods|Gadgets|Outfits|Things|Materials|Objects|Appliances|Resources|Solutions|Instruments|Components|Focus|Units} And Construction Throughout Austin - -Minoxidil Has Left Without Any Goodbye kinase 1 (CK1e)-dependent DVL2 phosphorylation on
serine 143 (Fig. 4c) that promotes DVL2 interaction with Polo-
like kinase-1 (PLK1), which subsequently activates the HEF-1/
PTEN´┐ŻCDsh interaction regulates morphogenetic processes. Aurora kinaseA/HDAC6pathway,triggeringtubulindeacetyla-
DVLisaparticularlyintriguingPTENpartner,asitiscrucialfor tion and cilia disassembly35,36.

These observations prompted us
ciliaformationinbothXenopusandmice. Inparticular, similar totestwhetherPTENcanregulatephospho-DVL2levels.Firstwe
towards the results of PTEN knockdown described over, reduction of compared the levels of phosphorylated serine 143 (pS143) on
DVL influences apical docking of BBs and overexpression Gear|Products|Items|Tools|Supplies|Devices|Machines|Accessories|Stuff|Goods|Gadgets|Outfits|Things|Materials|Objects|Appliances|Resources|Solutions|Instruments|Components|Focus|Units} And Release Throughout Vegas -Regorafenib Actually Leaves Without Any Kind Regards of the DVL2 in hTERT-RPE1 cells during the formation of cilia in


Report
DOI:10.1038/ncomms9390
OPEN
Received26Feb2015|Accepted17Aug2015|Published24Sep2015
Highlymultiplexedimagingofsinglecellsusinga
high-throughputcyclicimmuno?uorescence
process
Jia-RenLin1,MohammadFallahi-Sichani2&PeterK.

Sorger1,2
Single-cellanalysisrevealsaspectsofcellularphysiologynotevidentfrompopulation-based
studies, particularly in the case of highly multiplexed methods such as mass cytometry
(CyTOF) able to correlate the amounts of multiple signalling, differentiation and cell fate
markers.Immuno?uorescence(IF)microscopyaddsinformationoncellmorphologyandthe
microenvironmentthatarenotobtainedusing?ow-basedtechniques,butthemultiplicityof
conventionalIFislimited.Thishasmotivateddevelopmentofimagingmethodsthatrequire
specialized instrumentation, exotic reagents or proprietary protocols that are dif?cult to
reproduce in most laboratories. Here we report a public-domain technique for achieving
higher multiplicity single-cell IF using cyclic immuno?uorescence (CycIF), a simple and
versatile procedure in which four-colour staining alternates with chemical