Alimonti_Supplementary Figure 1. Pten +/- Pten + Pten. Pten hy. β-actin. Pten - wt hy/+ +/- wt hy/+ +/- Pten. Pten. Relative Protein level (% )
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1 Alimonti_Supplementry Figure 1 hy Neo Proe 5 Proe hy/ hy/ / Neo β-tin d Reltive Protein level (% ) hy/ /- Reltive Gene Expr. (% ) hy/ /- Supplementry Figure 1 Chrteriztion of hy/ mouse emryoni firolsts (MEFs). () Shemti representtion the genomi onstrut of the hypomorphi (hy) llele hroring the neomyin ssette, nd the wild-type (), nd null ( ) lleles. The genomi sequene is depited s white line, with gry oxes representing exons 3, 4, 5 nd 6. The retngulr ox represent the Neo resistne the gry tringles represent the loxp sites. The genomi frgment (proe 6.1), used s proe for Southern lot nlysis is shown s red r. () Western lot nlysis of, hy/ nd /- MEFs showing sutle redution of protein levels in hy/ MEFs when ompred with the nd /- ells. Arevition ode used is: hy/ = mie hving hypomorphi nd llele; /- = mie hving nd null llele () Quntifition of protein level from, hy/ nd /- littermte MEFs. Error rs show S.D. from three independent experiments. (d) Quntifition of mrna levels in, hy/ nd /- MEFs. Error rs show S.D. from three independent experiments. Nture Genetis: doi:1.138/ng.556
2 Alimonti _Supplementry Figure S2 (n=3) hy/ (n=32) / (n=24) Lymph-node Averge dimeter (inh) Perentge of mie ffeted with Lymphodenopthy n= m n=4 hy/ / >22 p=.2 Time (months) p=.8 d Spleen.35 Averge weigth(gr) hy/ /- n=5 n=4.3 n= hy/ /- 1 m n=3 Spleen Lymph-node hy/ 4X.15 4X.1 /-.5 hy/ /- 4X p=.4 p=.1 4X hy/ e liver 4X kidney 4X Supplementry Figure 2 Averge size of lymph nodes nd spleen in, hy/ nd /- mie. () Lymphodenopthy disese-free survivl in the, hy/ nd /- popultion; n= numer of nimls nlyzed. (-) Anlysis nd representtive imges of the verge size of the lymph nodes () nd spleen () of the, hy/ nd /- t the time of detetion of the utoimmune disorder. n= numer of nimls nlyzed. p= sttistil signifine. Error rs show S.D. (d) Hemtoxylin nd eosin (H&E) nlysis of spleens nd lymph-nodes showing lymphosplenomegly in hy/ nd /- mie. Inset: H&E stining of spleen nd lymph-node from wild-type mouse, showing spleen with well-defined white nd red pulp nd lymph-node with well-delineted mirontomil fetures. Note the hypertive white pulp nd the prtil effement of the spleen histology of the mutnt mie. Note lso the hyper retive germinl enters with the prtil effement of the norml ortiomedullry oundries of the hy/ nd /- lymph nodes. (e) H&E nlysis shows inflmmtory infiltrtions in the liver nd kidney of the hy/ mie. Nture Genetis: doi:1.138/ng.556
3 Alimonti_Supplementry Figure 3 hy/ /- Uterine dysplsi 2X 2X Lung Cner Intestinl Polyp hy/ 2X 2X Supplementry Figure 3 hy/ mie develop tumours of different histology fter long lteny. () Representtive H&E imges of hy/ nd /- uterus showing signs of uterine dysplsi. () (Left imge) H&E stining of multi-fol lung denorinom in hy/ femle mouse. (Right imge) Intestinl polyp from 16 month old hy/ mle (see lso Tle 1). Nture Genetis: doi:1.138/ng.556
4 Alimonti_Supplementry Figure 4 MAMMARY GLANDS lox PCR UTERUS PCR MAMMARY GLANDS UTERUS d 7k MAMMARY TUMORS e IHC in mmmry glnds 5k 7k /- Uterus 144 yy8 19 hy/- 4X 4X hy/ 5k /- 4X /- 4X hy/- 7k Uterus 5k 12 yy4 yy7 hy/- Supplementry Figure 4 hypomorphi mie develop mmmry nd uterine tumours in sene of LOH. () Sheme illustrting the PCR strtegy nd the PCR primers dopted for the LOH nlysis in the hy/ mie. () PCR nlysis of norml tissue, mmmry nd uterine tumours from, hy/ nd /- mie using the lox () nd () primers. Note tht three tumours for eh genotype were nlyzed for the LOH nlysis. (d) Southern lot nlysis of norml tissue, mmmry nd uterine tumours from, /- nd hy/- mie showing the sene of LOH in these tumours. Four tumours for eh genotype were nlyzed. (e) Representtive imges of immunohistohemistry nlysis for in the sme tumours s in (). Note tht tissues from oth hy/ nd /- mmmry epithelium show immunoretivity for. Nture Genetis: doi:1.138/ng.556
5 Alimonti_Supplementry Figure 5 hy/ Mmmry tissue H&E 1 1X 1X 1X β-tin Reltive Protein level (% ) 1X pakt 1X.5 / hy/ /- / hy/ / X.8 Supplementry Figure 5 Anlysis of pre-neoplsti mmmry glnds in ontrol nd hy/mie. () H&E nd immunohistohemistry nlysis for nd pakt in 2 months old nd hy/ virgin littermte femle mie. () (Upper pnel) Western lot nlysis for nd quntifition of protein level (Lower pnel) in two month old ontrol nd hy/ mmmry glnds showing the presene of protein levels in ll the tissues nlyzed. Error rs show S.D. from three independent experiments. Nture Genetis: doi:1.138/ng.556
6 Alimonti _Supplementry Figure d3 d2 sicontrol si Trnsripts (Reltive to Hprt) β-tin SiControl Si (d3) Reltive ell numer (OD 595 nm) 1,6 1,4 1,2 1,8,6,4,2 48 h p=.3 SiControl Si (d3) Supplementry Figure 6 A sutle down regultion of level y sirna promotes hyper-prolifertion in MMECs derived from mie of C57/BL6 pure geneti kground. (Left pnel) Down regultion of protein nd gene expression levels y seril dilutions of sirna in MMECs. (Right pnel) Prolifertion of MMECs derived from C57/BL6 mie, fter trnsfetion with the sirna dilution numer 3 (d3) in MMECs. Error rs show S.D. from three independent experiments. p= sttistil signifine. Nture Genetis: doi:1.138/ng.556
7 Alimonti _Supplementry Figure rest tumors norml rest GNF2_CDC2 GNF2_CSK1B Brest tumors PTEN Brest tumors Norml Brest Norml Brest 1,32 PTEN expression in rest tumors reltive to norml smples -1.5 Controls High PTEN expression,99,66,33 GSM85485 GSM85493 GSM85482 GSM85481 GSM8559 GSM85479 GSM85491 GSM85494 GSM85483 GSM85489 GSM85475 GSM85478 GSM85473 GSM85474 GSM85496 GSM85512 GSM8551 GSM85511 GSM8552 GSM8554 GSM85487 GSM85484 GSM8557 GSM85476 GSM8555 GSM85492 GSM85499 GSM8549 GSM85486 GSM85488 GSM8551 GSM8558 GSM855 GSM85497 GSM8548 GSM8556 GSM85477 GSM85498 GSM85495 GSM8553 GSM85514 GSM85515 GSM85513 GSM85517 GSM85516 GSM85519 GSM Ptients GNF2_CDC2 Brest tumors Norml Brest -4 4 PTEN gene expression (%) > 65 < 65-4 Numer of ptients Perentge of ptients GSM85498 (PTEN 7%) ID GSM85485 GSM85482 GSM855 GSM8548 GSM85498 GSM85495 PTEN mrna Reltive PTEN expression GSM85495 (PTEN 73%) PTEN IHC # * 4X # 2X GSM85482 (PTEN 22%) GSM85485 (PTEN 17%) # * 4X *Strom 4X # Norml epithelium Supplementry Figure 7 Chrteriztion of humn rest ner smples with deresed PTEN expression. () Gene expression profile from ontrol nd rest humn tumour smples with low/sent PTEN expression (upper pnel). Note tht these tumours show the sme up-regultion of gene sets enrihed in the hy/ MEFs. See lso Supplementry Fig. 8 for the GSEA nlysis of the enrihed gene sets in the ptients nlyzed nd jsp?olletion=cgn for the orresponding gene set pge. () Histogrm showing the distriution of PTEN mrna expression in norml mmmry tissue (lk rs) nd tumour speimens with sutle (PTEN mrna> 65%, ornge rs) nd pronouned (PTEN mrna <65%, lue rs) redution in PTEN mrna expression. The tle shows the numer nd perentge of ptient tumour smples exhiiting sutle (>65%) or more drmti (<65%) redution in PTEN mrna expression ompred to the verge PTEN expression in the norml mmmry tissue smples (n=7). () (Left pnel) Sore of PTEN immunoretivity in suset of ptients from () with vrying levels of PTEN mrna (showed s solute signl intensity vlues nd reltive expression ompred to norml mmmry tissue smples). (Right pnels) Representtive immunohistohemistry nlysis for PTEN in iopsies showing either sutle or profound dereses in PTEN mrna (orresponding PTEN mrna level reltive to norml mmmry tissue is indited in rkets) Nture Genetis: doi:1.138/ng.556
8 Alimonti_Supplementry Figure rest tumor d 4. norml rest e f g Supplementry Figure 8. Gene expression profile in ontrol nd ptients ffeted y invsive rest ners. (Upper pnels) Enrihment plot ssessed y GSEA for GNF2_BUB1(), GNF2_CDC2 (), GNF2_CDC2 (), GNF2_MCM4 (d), GNF2_CENPE (e), GNF2_CKS2 (f) nd GNF2_CKSIB (g) gene sets in tumours smples nd in the hy/ MEFs. Genes in the linege-speifi gene sets re mrked with vertil rs, nd the enrihment sore is shown in green. (Lower pnels) Mrker genes of the GNF2_BUB1(), GNF2_CDC2 (), GNF2_CDC2 (), GNF2_ MCM4 (d), GNF2_CENPE (e), GNF2_CKS2(f) nd GNF2_CKSIB (g) gene sets, in ptients ffeted y rest ner (with deresed gene expression for PTEN) ompred to ontrols, rnked y signl to noise rtio Nture Genetis: doi:1.138/ng.556
9 Supplementry Tle 1. Primer sequenes used for genotyping, exons muttionl nlysis nd sirna experiment. GENOTYPING Forwrd 5 >3 Reverse 5 >3 TGGGAAGAACCTAGCTTGGAGG TTCCATTTGTCACGTCCTGCAC ACTCTACCAGCCCAAGGCCCGG LPTEN1 TGTTTTTGACCAATTAAAGTAGGCTGTG AAAAGTTCCCCTGCTGATGATTTGT MUTATIONAL ANALYSIS exon Forwrd 5 >3 Reverse 5 >3 1 AGTCCAGAGCCATTTCCATC TCTAGAAATGCGCCCAGAAT 2 CTAGCGTGGGAAAAGCTAA CCAAAACACATCAAAATGCAA 3 CTGTTTTAGTCCTGTGCAGC CAGGAATCCAGTTCCTCAAA 4 AGCGCAGTGTTTTTACATGA TCACCAGGCAGTAAAAGAC 5 TGGAGTGAAGAGCACAATGC CACAAAGAGGGAGGAAGGAA 6 TTTGTCTCCCTCCTCCCTCT CACCAAAATTCATTGGGTTAGC 7 GAAGTCCTTACATGGGTTGG AAGGCTTTAAGCAAAAGGTCTG 8 CACAAGGTGTTTGCCTTCAC CAGAAAAGGAAAGGCTACGC 9 AAAAGCAGTGCCCTTCAGA CCTATAACTGCAATCTGACA sirna EXPERIMENT sirna ControlsiRNA AGACCAUAACCCACCACAG AAGGAGACGAGCAAGAGAA Nture Genetis: doi:1.138/ng.556
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doi: 1.138/nture862 humn hr. 21q MRPL39 murine Chr.16 Mrpl39 Dyrk1A Runx1 murine Chr. 17 ZNF295 Ets2 Znf295 murine Chr. 1 COL18A1 -/- lot: nti-dscr1 IgG hevy hin DSCR1 DSCR1 expression reltive to hevy
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oi:1.138/nture1138 Supplementl Figure 1 Inflmmtory Monoytes Host ells CCR2 CCL2 Disseminting Tumor Cells Metstsis Assoite Mrophges VEGF Extrvstion & Metstti Seeing Supplementl Figure 1 The t from this
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Copy Number ID2 MYCN ID2 MYCN. Copy Number MYCN DDX1 ID2 KIDINS220 MBOAT2 ID2
Copy Numer Copy Numer Copy Numer Copy Numer DIPG38 DIPG49 ID2 MYCN ID2 MYCN c DIPG01 d DIPG29 ID2 MYCN ID2 MYCN e STNG2 f MYCN DIPG01 Chr. 2 DIPG29 Chr. 1 MYCN DDX1 Chr. 2 ID2 KIDINS220 MBOAT2 ID2 Supplementry
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doi:.3/nture93 d 5 Rttlesnke DRG (reds) Rttlesnke TG (reds) c 3 TRPV1 other TRPs 1 1 3 Non-pit snke TG (reds) SFig. 1 5 5 3 other TRPs TRPV1 1 1 3 Non-pit snke DRG (reds) 5 Antomy of the pit orgn nd comprison
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Supplementary Information
Supplementry Informtion Cutneous immuno-surveillnce nd regultion of inflmmtion y group 2 innte lymphoid cells Ben Roediger, Ryn Kyle, Kwok Ho Yip, Nitl Sumri, Thoms V. Guy, Brin S. Kim, Andrew J. Mitchell,
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doi:1.138/nture1228 Totl Cell Numer (cells/μl of lood) 12 1 8 6 4 2 d Peripherl Blood 2 4 7 Time (d) fter nti-cd3 i.p. + TCRβ + IL17A + cells (%) 7 6 5 4 3 2 1 Totl Cell Numer (x1 3 ) 8 7 6 5 4 3 2 1 %
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X p -lu c ct ivi ty doi:.8/nture8 S CsA - THA + DAPI Merge FSK THA TUN Supplementry Figure : A. Ad-Xp luc ctivity in primry heptocytes exposed to FSK, THA, or TUN s indicted. Luciferse ctivity normlized
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DOI:.38/nc393 Nnog DAPI Nnog/DAPI c-jun DAPI c-jun/dapi c e Reltive expression to Gpdh mescs ( Feeder free) mescs ( Feeder) MEFs.5 MEFs ipscs ESCs..5 p=.24 p=.483 p=.22. JunB JunD Fos Fr Fr2 ATF2 ATF3
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% DAPI + Are Supplementry Figure 1 MOI 1 MOI.5 MOI.25 5 4 3 MOPC MOPC, LCMV reltive VL4 expression 2 1 d1 d2 d3 MOI.125 MOI.625 untreted Supplementry Figure 1: LCMV replictes in MOPC without ffecting cell
NappHS. rrna. transcript abundance. NappHS relative con W+W 0.8. nicotine [µg mg -1 FM]
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(A) W+OS 3 min 6 min con L S L S RNA loding control NppHS rrna (B) (C) 8 1 k NppHS reltive trnscript undnce 6 4.5 *** *** *** *** 3 k. + + + line 1 line (D) nicotine [µg mg -1 FM] 1..8.4. con W+W Supplementl
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doi:.38/nture277 d 25 25 2 Time from sound onset (ms) 25 25 2 Time from sound onset (ms) Firing rte (spikes/s) Firing rte (spikes/s).8.6..2 e f g h.8.6..2 Frtion of neurons Frtion of neurons N = 53 2 2
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Supplementary Figure 1
Roles of endoplsmic reticulum stress-medited poptosis in -polrized mcrophges during mycocteril infections Supplementry informtion Yun-Ji Lim, Min-Hee Yi, Ji-Ae Choi, Jung-hwn Lee, Ji-Ye Hn, Sung-Hee Jo,
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Chow KD CR HFD. Fed Fast Refed
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DOI: 10.1038/nc2824 Hcn4 Tx5 Mlc2 c Hcn4- ISH d Tx5- ISH e Mlc2-ISH Hcn4-ISH f e Tx5-ISH f -ISH Figure S1 Section in situ hyridistion nlysis of crescent stge mouse emryos (E7.5). () More nterior section
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regulates stem cells through Wnt/β-catenin signalling
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SUPPLEMEARY IFORMAIO doi:./nture correction to Supplementry Informtion Adenom-linked rrier defects nd microil products drive IL-/IL-7-medited tumour growth Sergei I. Grivennikov, Kepeng Wng, Dniel Mucid,
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SUPPLEMENTARY INFORMATION doi: 1.138/nnno.211.41 Sili nd titnium dioxide nnoprtiles use pregnny omplitions in mie Kohei Ymshit, Ysuo Yoshiok, Kzum Higshisk, Kzuy Mimur, Yuki Morishit, Mstoshi Nozki, Tokuyuki
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% cells forming Neurospheres 81 ± 6 % 0 % 2.6 ± 0.7 % 76 ± 8 % 0 % 3.4 ± 0.6 % 83 ± 5 % 0 % 2.4 ± 0.9 % 89 ± 5 % 3 ± 1.5 % Total 10, ± 6 % 0 %
Bo et l., Suppl. Tle 1 Supplementl Tle 1. Neurosphere formtion nd tumorigencity is enriched within the tumour cell popultions derived from humn primry glioms nd gliom xenogrfts. GBM smples or Gliom xenogrfts
Supplementary Figure S1
Supplementry Figure S1 d MAP2 GFAP e MAP2 GFAP GFAP c f Clindin GFAP Supplementry Figure S1. Neuronl deth nd ltered strocytes in the rin of n ffected child. Neuron specific MAP2 ntiody stining in the hippocmpus
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SUPPLEMENTARY INFORMATION doi:10.1038/nture11225 Numer of OTUs sed on 3% distnce Numer of 16s rrna-sed V2-V4 tg sequences LF MF PUFA Supplementry Figure 1. High-ft diets decrese the richness nd diversity
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residual wild type band (Supplementary Fig. 5c) that may be explained by the presence of
doi:1.138/nture9387 Supplementry Text RANK deletion in tumors nd Cre effects. Southern lotting of the tumors tht developed in RANK mm femles showed deletion of RANK, leit we oserved some residul wild type
* * * * * liver kidney ileum. Supplementary Fig.S1
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Supplementary Figure S1
Supplementry Figure S Connexin4 TroponinI Merge Plsm memrne Met Intrcellulr Met Supplementry Figure S H9c rt crdiomyolsts cell line. () Immunofluorescence of crdic mrkers: Connexin4 (green) nd TroponinI
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Control vector. HA-Elfn2. HA-Elfn1
Control vector c HA-Elfn2 HA- Control vector HA-Elfn2 HA- nti- nti-ha Hippocmpus (CA3) PV DAPI SOM DAPI Hippocmpus (dentte gyrus) PV DAPI SOM DAPI Supplementry Figure 1. Specificity of the nti- ntiody
2018 American Diabetes Association. Published online at
Supplementry Figure S1. Ft-1 mice exhibit reduced diposity when fed n HFHS diet. WT nd ft-1 mice were fed either control or n HFHS diet for 18 weeks. A: Representtive photogrphs for side-by-side comprison