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15
15.4
14.6
15
15.4
DNA content (log AU)
DNA content (log AU)
DNA information (log AU)
d
e
viSNE maps coloured by signal intensities
Ki-67
p-Rb
p-S6
S235/236
p-S6
S240/244
p-S6S240/244
Ki-67
Large
Substantial
Population truly 2: Cycling
10
8
1.2
0.8
0.4
0
6
4
2
0
Population 1: Noncycling
Lower
Lower
0
0.2
0.4
0.6
0.8
1
Wanderlust axis
Figure3|Multivariatesingle-cellanalysisofBRAFV600ECOLO858melanomacellsusingCycIF.(a)Highlysimpli?edschematicshowingcrosstalk
betweenMAPKandPI3K/Akt/mTORpathwaysatthelevelofS6ribosomalproteinphosphorylation.(b)Unsupervisedclusteringofsingle-cell8-channel
CycIFdataincellstreatedwithDMSOor0.1mMvemurafenibfor24h.ThehierarchicalclusteringwasperformedinMatlabusingEuclideandistance
metricsandaveragelinkage.

(c)Scatterplotscomparingthreechannelsatatime(p-Rb,Hoechst,p21andKi-67)todeterminethedistributionofG0,G1/S
andG2cellcyclestates.(d)viSNE-generatedtwo-dimensionalprojectionsofthesingle-cell8-channelCycIFdata.Single-celldataareplottedinviSNEaxes
andcolouredbasedonKi-67(left)andpS6S240/244(ideal)signalintensities.(e)Wanderlusttraceshowingsignalintensitiesforp-RbS807/811,pS6S235/236
,
Ki-67andpS6S240/244acrossa?ve-pointvemurafenibdose�Cresponse(0�C1mM)dataset.Thecolour-boxonthebottomrepresentsthecelldensitieswhile

thecolourscaleisshowedontheside.
NATURE http://www.selleckchem.com/products/AG-014699.html COMMUNICATIONS |6:8390|DOI: 10.1038/ncomms9390 |www.nature.com/naturecommunications
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&2015MacmillanPublishersLimited.Allrightsreserved.




Inhibition of hedgehog signaling ameliorates hepatic inflammation in
mice with nonalcoholic fatty liver disease (NAFLD)
Hyunjoo Kwon1*, Kyoungsub Song1*, Chang Han1, Weina Chen1, Ying Wang1,2, Srikanta
Dash6, Kyu Daptomycin Lim3 and Tong Wu1
1Department of Pathology and Laboratory Medicine, Tulane University School of
Medicine, 1430 Tulane Avenue SL