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1 2 weeks high holesterol diet 2 weeks high holesterol diet 2 weeks high holesterol diet 2 μm Mrophges Crystls Hoehst μm Mrophges Crystls Hoehst Hoehst Crystls Mrophges 2 μm 2 μm Supplementry Fig. 1: Erly orti sinus therosleroti lesions ontin rystl-loded mrophges. -, Comined onfol fluoresene nd refletion mirosopy of erly orti sinus therosleroti lesions of Apo-E-KO mie fed high holesterol diet for 2 weeks. Lser refletion of holesterol rystls (red), immunofluoresene of mrophges (green) identified y MoM-2 nd nulei (lue) ounterstined with Hoehst dye. Note: In erly lesions individul suendothelil rystl-positive mrophges n e seen () nd grnulom-like lesions of few tens of mrophges enirling holesterol rystl depositions re visile (). Additionlly, holesterol-rystl positive mrophges n e found in leflets of the hert vlves (). Representtive imges from seril setions of groups of 3 mie re shown. 1
2 3 μm Crystls Crystls Hoehst Dendriti ells Crystls 2 μm Crystls Hoehst Dendriti ells Crystls 2 Crystls Hoehst Dendriti ells Mrophges Crystls Crystls Hoehst Neutrophils Mrophges Supplementry Fig. 2: Erly therosleroti lesions ontin rystl-loded mrophges t different sites of orte. -, Comined onfol fluoresene nd refletion mirosopy of erly orti sinus therosleroti lesions of Apo-E-KO mie fed high holesterol diet for 4 weeks. Smples were stined for () dendriti ells (CD11, red), () mrophges (MoM-2, red) nd dendriti ells (CD11, green) or () mrophges (MoM-2, red) nd neutrophils (nti-neutrophil, green). Nulei were ounterstined with Hoehst dye. Representtive imges from seril setions of groups of 3 mie re shown. 2
3 Tuni dventiti Tuni medi Atherosleroti plque Tuni intim Arteril lumen 1 μm Mrophges (CD68) Smooth musle ells (phlloidin) Nulei (Hoehst) Crystl (refletion) 3 μm 2 μm Supplementry Fig. 3: Humn oronry rtery therosleroti lesions ontin rystls suendothelilly nd in neroti ores. Comined onfol fluoresene nd refletion mirosopy of humn oronry rtery therosleroti lesion. Overview imge ws otined without refletion mirosopy, while mgnifitions of oxed res were otined using omined immunofluoresene nd refletion detetion. Tissue ws stined for mrophges (CD68, red), smooth musle ells (phlloidin, light lue) nd nulei (Hoehst dye, drk lue). A representtive setion of one ptient is shown. 3
4 3 μm Mrophges Crystls Hoehst Mrophges Crystls Hoehst Trihrome stin 2 μm μm 2 μm 2 μm Supplementry Fig. 4: Humn oronry rtery lesion ontining lrge holesterol rystls -, Comined onfol fluoresene nd refletion mirosopy of humn oronry rtery therosleroti lesion. Lesion ws stined with Msson s trihrome stin (). Note: Lrge holesterol rystl lefts re visile, ut smll holesterol rystls re not pprent. (, ) Lesions were stined for mrophges (CD68, red). Note: Lrge holesterol rystls re found extrellulrly () nd smll rystls re found intrellulrly (). Smple imged t 37 C using wrmstge. Representtive setions of two ptients (, ) (different from ptient shown in Supplementry Fig. 3) re shown. 4
5 6 3 Syntheti holesterol + MCD MCD Drkfield mirosopy IL18 (pg/ml) Cholesterol rystls (μg/ml) MDP-priming 2 1 Cholesterol rystls (µg/ml) Nigeriin 2 1 iedap-priming 16 Cholesterol (μg/ml) d Cholesterol rystls LDL priming (µg/ml) ATP 1 2 Cholesterol rystls ATP LPS priming Supplementry Fig. 5: Cholesterol rystl-medited IL-1β relese in primry murine onemrrow ells is dependent of the NLRP3 inflmmsome., IL-18 ELISA of superntnts from LPS-primed wild-type mrophges stimulted with holesterol rystls or nigeriin., IL-1β ELISA of superntnts from LPS-primed one mrrow derived DCs stimulted with syntheti holesterol rystls with or without methylylodextrin (MCD). Equivlent doses of MCD were diluted into medi of LPS-primed mrophges. Note: Upon dilution in queous ulture medi, smll holesterol rystls fll out of solution y dilution of the holesterol-soluilizing gent MCD (, right)., d, IL-1β ELISA of superntnts from () MDP- or iedap- nd (d) etylted LDL- or LPS-primed mouse mrophges, stimulted with holesterol rystls, ATP or left untreted. Mens with s.e.m. from one out of two (,, d) three () representtive experiments re shown. 5
6 2 16 No Inhiitor Ltrunulin A Overly fluoresene DiD DiI holesterol rystlsholesterol rystls IL-1β in pg/ml Ex: 488 nm; Em: mn e Cell numer Cell numer 2 1 dadt Cholesterol rystls (μg/ml) Monoytes.63 DiI holesterol rystls 2% mdcs.63 DiI holesterol rystls Overly + trnsmitted light pdcs.63 DiI holesterol rystls μg/ml 2 μg/ml μg/ml 1 μg/ml 4% 6% 9% Aridine ornge fr red fluoresene (lysosome) 18% Overly + refletion f Lymphoytes DiI holesterol rystls d 1 Bfilomyin (nm) none Cholesterol rystls (μg/ml) Supplementry Fig. 6: Fluoresent holesterol rystls re phgoytosed y monoytes, mrophges nd dendriti ells., IL-1β ELISA of superntnts from LPS-primed mouse mrophges stimulted with holesterol rystls or dadt in the presene or sene of Ltrunulin A., A mixture of unleled, DiI-leled (green) or DiD-leled (red) holesterol rystls ws inuted for 24 h nd susequently imged y omintion of refletion (unleled, white) or fluoresene onfol imging. Note: Dyes ound to the rystls do not exhnge nd non-leled rystls do not quire dye., PBMCs were inuted with DiI-leled holesterol rystls in the sene or presene of inresing mount of ytohlsin D. Cells were stined with surfe mrkers to distinguish ell popultions nd nlyzed y flow ytometry. The men fluoresene intensity of DiI-holesterol rystl uptke is shown for the different ell popultions. d, LPS-primed humn PBMCs were left untreted or treted with ytohlsin D nd susequently inuted with 2 μg/ml holesterol rystls. IL-1β ws determined in superntnts y ELISA. e, Murine mrophges stimulted with holesterol rystls for 6h, stined with ridine ornge nd nlyzed y FACS. f, IL-1β ELISA of superntnts from LPSprimed, murine mrophges stimulted with holesterol rystls in the presene or sene of filomyin A1. Dt re representtive of one experiment repeted twie (-d) or three times (e, f). 6
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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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SUPPLEMENTARY INFORMATION doi:.8/nture89 4 4 Ilr -/- Ilr -/- Ilr -/- Cspse- -/- As -/- Nlrp -/- Il8 -/- Ilr -/- Supplementl figure. Inresed severity of NASH in inflmmsome-defiient mie, ut not in Ilr-defiient
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doi:10.1038/nture09663 Scrmle shnlrp3 shcsp1 IL-1β (p17) IL-1β (pg/ml) 2000 1500 1000 500 Wt Nlrp3-/- Ipf-/- 0 APDC IL-1β (p17) Supplementl Figure 1. Mitochondril ROS cn trigger NLRP3 inflmmsome ctivtion,
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DOI: 1.138/n358 TLR2 nd MyD88 expression in murine mmmry epithelil supopultions. CD24 min plus MRU Myo-epithelil Luminl progenitor (CD61 pos ) Mture luminl (CD61 neg ) CD49f CD61 Reltive expression Krt5
Supplementary Figure S1
Supplementry Figure S1 - UTR m - 3HA - 2-1 hgh - 1 Uiquitin *! *! lk distl promoter m K3R/ K121R-3HA UTR hgh founder lines - HA - - founder lines TG- E1 L A2 B1 F9 G6 H4 H6 B C D2 G1 H3 J2 L - 7 IP: lk
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DOI: 1.13/n7 Reltive Pprg mrna 3 1 1 Time (weeks) Interspulr Inguinl Epididyml Reltive undne..1.5. - 5 5-51 51-1 1-7 7 - - 1 1-1 Lipid droplet size ( m ) 1-3 3 - - - 1 1-1 1-1 1-175 175-3 3-31 31-5 >5
Silica crystals and aluminum salts activate the NALP3 inflammasome through phagosomal destabilization
8 Nture Pulishing Group http://www.nture.om/ntureimmunology rystls nd luminum slts tivte the NALP3 inflmmsome through phgosoml destiliztion Veit Hornung 1,, Frnz Buernfeind,, Annett Hlle 1, Eivind O Smstd
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% ells with ili (mrke y A-Tu) Reltive Luiferse % ells with ili (mrke y Arl13) % ells with ili DOI: 1.138/n2259 A-Tuulin Hoehst % Cilite Non-ilite -Serum 9% 8% 7% 1 6% % 4% +Serum 1 3% 2% 1% % Serum: -
(% of adherent cells) *** PBL firm adhesion. Frequency (% ) 4 1 L 2 CXCR3 DP-2
Chemotxis (% of dded ells) PBL totl dhesion (N ells/mm 2 /1.1 6 PBL) Frequeny (% ) PBL firm dhesion Supplementry Figure 1 4 4 3 3 2 2 1.1-4 1-3 1.1.2. 1 1 8 6 4 2 Adiponetin ( g/ml) - + Adiponetin ( g/ml)
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Xio, et l., Supplementry Figure 1 IL-6 (ng/ml) TNF-α (pg/ml) 16 12 8 4 1,4 1,2 1, 8 6 4 2 med Cl / Pm3CSK4 zymosn curdln Poly (I:C) LPS flgelin MALP-2 imiquimod R848 CpG TNF-α (pg/ml) IL-6 (ng/ml) 2 1.6
Cos7 (3TP) (K): TGFβ1(h): (K)
IP#2: IP#1: Totl Lystes luiferse tivity (K): 6-4 - (K): luiferse tivity luiferse tivity (K): 2 1 RL-: - + + + + + Sm4-3F: + - + + + + MYC-Sm3: - - - - + + TβRI-HA(T204D): - - - + - + α-ha Luiferse Ativity
Supplementary Information
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Alimonti_Supplementary Figure 1. Pten +/- Pten + Pten. Pten hy. β-actin. Pten - wt hy/+ +/- wt hy/+ +/- Pten. Pten. Relative Protein level (% )
Alimonti_Supplementry Figure 1 hy 3 4 5 3 Neo 4 5 5 Proe 5 Proe hy/ hy/ /- - 3 6 Neo β-tin d Reltive Protein level (% ) 15 1 5 hy/ /- Reltive Gene Expr. (% ) 15 1 5 hy/ /- Supplementry Figure 1 Chrteriztion
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RESEARCH ARTICLE. Supplemental Figure 5
11.5 2 2 11. RESEARCH ARTICLE RBC ( 1 12 /L) 1.5 1. 9.5 PLT ( 1 9 /L) 1 16 14 HGB (g/l) 19 1 17 16 9. 12 4 4 46 Cellulr & Moleulr Immunology dvne online pulition, PCV (%) 44 MCV (fl) 46 44 ; doi:1.13/mi.214.16
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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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DOI:.3/n95 Thymus Kiney (kd) TA T7 T TA T7 T Hert TA T7 T: +Dox Cylin B (kd) Thymus Kiney Hert TA T5 T TA T5 T TA T5 T: +Dox Cylin B Poneu S Poneu S CnB T7 CnB T Thymus (kd) + Liver Colon + + (kd) Thymus
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Prentl doi:.8/nture57 Figure S HPMECs LM Cells Cell lines VEGF (ng/ml) Prentl 7. +/-. LM 7. +/-.99 LM 7. +/-.99 Fold COX induction 5 VEGF: - + + + Bevcizum: - - 5 (µg/ml) Reltive MMP LM mock COX MMP LM+
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doi: 1.138/nture6188 SUPPLEMENTARY INFORMATION Supplementry Figure 1 c CFU-F colonies per 1 5 stroml cells 14 12 1 8 6 4 2 Mtrigel plug Neg. MCF7/Rs MDA-MB-231 * * MCF7/Rs-Lung MDA-MB-231-Lung MCF7/Rs-Kidney
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doi:1.138/nture1794 BR EPFs BRI1? ERECTA TMM BSKs YDA PP2A BSU1 BIN2 pbzr1/2 BZR1/2 MKK4/5/7/9 MPK3/6 SPCH Cell growth Stomtl production Supplementry Figure 1. The model of BR nd stomtl signling pthwys.
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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 %
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AMPK maintains energy homeostasis and survival in cancer cells via. regulating p38/pgc-1α-mediated mitochondrial biogenesis
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NC bp. b 1481 bp
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Supplementary figure 1
Supplementry figure 1 Dy 8 post LCMV infection Vsculr Assoc. Prenchym Dy 3 post LCMV infection 1 5 6.7.29 1 4 1 3 1 2 88.9 4.16 1 2 1 3 1 4 1 5 1 5 1.59 5.97 1 4 1 3 1 2 21.4 71 1 2 1 3 1 4 1 5 1 5.59.22
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~ 5 % ltion > 99 % ltion 25 2 15 5 Gly yemi (mmol/l) 3 7 15 3 Time fter -ell ltion (dys) Survivl (%) & ~ 5 % ltion 75 > 99% ltion 5 25 25 5 (dys) 7.4 d 1.25 lood ph 73 7.3 7.2 7.1 7. 7d ody weight h nge
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DOI:./n BJ RAS:ER Herrnz et l Supplementry Figure HFFF RAS:ER.. mrna Expression..... ILα ILβ IL IL CCL INH VEGF mrna Expression..... ILα ILβ IL IL CCL INH VEGF + OHT Torin NVP-BEZ + OHT shmtor. shmtor.
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oi:1.138/nture1134 CS+ CS- MCH 3 OCT OCT 3 MCH CS- CS+ OCT MCH 3 MCH OCT 3 OCT vs MCH OCT vs MCH ppetitive memory (PI) A 1-1 Unpire onitioning DDC-GAL4/UAS-Trp UAS-Trp/+ -2 MCH OCT OCT MCH sugr OCT MCH