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Supplementary Table S1. Clinicopathological characteristics of HCC patients Discovery Set 1 Discovery Set 2 Validation Set 1 Validation Set 2 Total no. of patients 10 10 20 44 Age, years Mean (SD) 66.1 (5.9) 68 (8.9) 61.3 (10.9) 64.8 (9.9) Sex, no. of patients (%) Male 10 9 16 (80) 35 (79.5) Female 0 1 4 (20) 9 (20.5) Hepatitis virus infection, no. of patients (%) No 1 1 2 (10) 9 (20.5) B 2 5 7 (35) 12 (27.3) C 6 3 11 (55) 20 (45.5) B+C 1 1 0 3 (6.8) Background liver, no. of patients (%) Normal 0 0 1 (5) 5 (11.4) Chronic hepatitis (CH) 7 3 8 (40) 23 (52.3) Liver cirrhosis (LC) 2 4 8 (40) 8 (18.2) CH + LC 0 0 1 (5) 0 Precirrhosis 1 3 2 (10) 8 (18.2) Tumor size, cm3 mean (SD) 68.3 (42.0) 120.9 (218.1) 87.8 (111.9) 176.4 (256.7) Histology, no. of patients (%) Well 1 0 1 (5) 5 (11.4) Well to moderately 0 0 4 (20) 0 Moderately 5 10 8 (40) 27 (61.4) Moderately to poorly 1 0 1 (5) 0 Poorly 3 0 6 (30) 12 (27.3) Intrahepatic metastases, no. of patients (%) 0 7 7 17 (85) 30 (68.2) 1 1 1 2 (10) 14 (31.8) 2 2 2 0 0 3 0 0 1 (5) 0 Portal vein involvement, no. of patients (%) 0 4 1 10 (50) 17 (38.6) 1 5 8 7 (35) 24 (54.5) 2 0 1 0 1 (2.3) 3 1 0 3 (15) 2 (4.5) Supplementary Table S2. List of genes up-regulated in HCC (Discovery Set 1) Transcript Fold RefSeq Gene symbol P-value Gene description Cluster ID change 2337716 NM_006252 PRKAA2 6.71E-04 4.42 Protein kinase, AMP-activated, alpha 2 catalytic subunit 2379863 NM_016343 CENPF|TUBD1|P 1.05E-04 3.22 Centromere protein F, 350/400ka (mitosin) | tubulin, delta AK1 1 | p21/Cdc42/Rac1-activated kinase 1 (STE20 homolog, yeast) 2427007 NM_002959 SORT1|PSRC1|M 1.24E-04 3.14 Sortilin 1 | proline/serine-rich coiled-coil 1 | myosin YBPHL binding protein H-like 2441386 NM_003617 RGS5|UPK1B 5.06E-06 3.27 Regulator of G-protein signalling 5 | uroplakin 1B 2449559 NM_018136 ASPM 4.37E-05 3.03 Asp (abnormal spindle) homolog, microcephaly associated (Drosophila) 2469252 NM_001034 RRM2 6.68E-06 3.36 Ribonucleotide reductase M2 polypeptide 2544201 NM_004881|NM_147 TP53I3|PFN4 7.69E-06 3.67 Tumor protein p53 inducible protein 3 | profilin family, 184 member 4 2784113 NM_001237 CCNA2 4.01E-04 3.43 Cyclin A2 2813414 NM_031966 CCNB1 5.02E-06 3.53 Cyclin B1 2899102 NM_003531 HIST1H3C 5.21E-05 4.94 Histone cluster 1, H3c 2946215 NM_003537 HIST1H3B 3.73E-06 4.99 Histone cluster 1, H3b 2947077 NM_003533 HIST1H3I 2.49E-05 3.61 Histone cluster 1, H3i 2997376 NM_018685 ANLN 1.43E-05 3.39 Anillin, actin binding protein 3025433 NM_020299 AKR1B10|LOC34 4.50E-04 18.03 Aldo-keto reductase family 1, member B10 (aldose 0888|LOC441282 reductase) | similar to aldo-keto reductase family 1, member B10 3260586 NM_005063 SCD|LOC645313| 8.76E-05 3.43 Stearoyl-CoA desaturase (delta-9-desaturase) | similar to LOC651109 Acyl-CoA desaturase (Stearoyl-CoA desaturase) (Fatty acid desaturase) (Delta(9)-desaturase) 3329343 NM_002391|NM_001 MDK 4.38E-04 3.90 Midkine (neurite growth-promoting factor 2) 012334|NM_0010123 33 3375545 NM_013402 FADS1|FADS3 2.74E-04 3.34 Fatty acid desaturase 1 | fatty acid desaturase 3 3590388 NM_018454|NM_016 NUSAP1 1.62E-05 3.09 Nucleolar and spindle associated protein 1 359 3756193 NM_001067 TOP2A 7.29E-06 4.63 Topoisomerase (DNA) II alpha 170kDa 3881443 NM_012112 TPX2|TINAGL1 2.10E-06 3.11 TPX2, microtubule-associated, homolog (Xenopus laevis) | tubulointerstitial nephritis antigen-like 1 3639031 NM_199414|NM_199 PRC1 8.32E-06 3.17 Protein regulator of cytokinesis 1 413|NM_003981 Supplementary Table S3. List of genes down-regulated in HCC (Discovery Set 1) Transcript Fold RefSeq Gene symbol P-value Gene description Cluster ID change 2331679 NM_032793 MFSD2|LOC731362 1.37E-05 3.98 Major facilitator superfamily domain containing 2 | similar to ribosomal protein S2 2337786 NM_000562 C8A 6.13E-04 3.77 Complement component 8, alpha polypeptide 2377332 NM_000651|NM_000 CR1|LOC653907 3.69E-08 3.32 Complement component (3b/4b) receptor 1 (Knops blood 573 group) | similar to complement component (3b/4b) receptor 1 isoform F precursor 2388085 NM_003679 KMO 2.13E-04 3.96 Kynurenine 3-monooxygenase (kynurenine 3-hydroxylase) 2403080 NM_003665|NM_173 MAP3K6|FCN3 4.00E-09 18.47 Mitogen-activated protein kinase kinase kinase 6 | ficolin 452 (collagen/fibrinogen domain containing) 3 (Hakata antigen) 2439138 NM_005894 CD5L 1.52E-06 12.18 CD5 molecule-like 2443120 NM_001937 DPT 4.15E-07 5.63 Dermatopontin 2475911 NM_014600 EHD3 3.24E-09 3.30 EH-domain containing 3 2502686 NM_006770 MARCO 8.19E-10 6.79 Macrophage receptor with collagenous structure 2547231 NM_000348 SRD5A2 7.70E-05 4.20 Steroid-5-alpha-reductase, alpha polypeptide 2 (3-oxo-5 alpha-steroid delta 4-dehydrogenase alpha 2) 2678298 NM_004944 DNASE1L3|ACTB 5.72E-07 9.65 Deoxyribonuclease I-like 3 | actin, beta 2693620 NM_144639 UROC1 2.30E-06 6.38 Urocanase domain containing 1 2716124 NM_001528 HGFAC 9.54E-07 4.84 HGF activator 2729852 NM_005953 MT2A|MT1M|LOC4 1.53E-04 3.38 Metallothionein 2A | metallothionein 1M | hypothetical 41019|NUTF2 gene supported by X97260; BC070289 | nuclear transport factor 2 2745899 NM_022475 HHIP 7.34E-08 3.97 Hedgehog interacting protein 2772566 NM_144646 IGJ 4.02E-04 7.76 Immunoglobulin J polypeptide, linker protein for immunoglobulin alpha and mu polypeptides 2773434 NM_002089 CXCL2 1.94E-05 5.90 Chemokine (C-X-C motif) ligand 2 2775259 NM_152545 RASGEF1B|ALK|R 6.25E-05 3.25 RasGEF domain family, member 1B | anaplastic BM41 lymphoma kinase (Ki-1) | RNA binding motif protein 41 2775390 MOP-1 8.90E-04 3.53 2775909 NM_016619 PLAC8 7.54E-11 13.74 Placenta-specific 8 2792127 NM_000909 NPY1R|LOC729743| 6.08E-09 3.06 Neuropeptide Y receptor Y1 | similar to neuropeptide Y LOC731120 receptor Y1 2794584 NM_005277|NM_201 GPM6A|MPDU1 9.68E-09 3.47 Glycoprotein M6A | mannose-P-dolichol utilization defect 591|NM_201592 1 2807716 NM_000587 C7 2.60E-05 11.43 Complement component 7 2816536 NM_001882 CRHBP 3.45E-09 13.85 Corticotropin releasing hormone binding protein 2842624 NM_017675 PCLKC 6.73E-04 4.88 Protocadherin LKC 2853055 NM_031900 AGXT2 4.55E-04 3.61 Alanine-glyoxylate aminotransferase 2 2854092 NM_002310 LIFR 2.95E-07 7.18 Leukemia inhibitory factor receptor alpha 2854409 NM_001737 C9|ASTN2 7.35E-05 16.46 Complement component 9 | astrotactin 2 2876608 NM_004887 CXCL14 8.13E-08 4.89 Chemokine (C-X-C motif) ligand 14 2883283 NM_138379 TIMD4 3.38E-08 4.36 T-cell immunoglobulin and mucin domain containing 4 2907513 NM_018960 GNMT 1.61E-04 6.98 Glycine N-methyltransferase 2955761 NM_016593 CYP39A1 6.59E-04 3.97 Cytochrome P450, family 39, subfamily A, polypeptide 1 2975257 NM_022568|NM_170 ALDH8A1|HBS1L 7.13E-04 3.41 Aldehyde dehydrogenase 8 family, member A1 | 771 HBS1-like (S. cerevisiae) 2982630 NM_024492|NM_145 LPAL2|LOC732058| 5.06E-05 3.86 Lipoprotein, Lp(a)-like 2 | similar to Apolipoprotein(a) 727 PLGLB2 precursor (Apo(a)) (Lp(a)) | plasminogen-like B2 2982730 NM_005577 LPA|LOC732390|L 8.05E-06 7.98 Llipoprotein, Lp(a) | similar to Apolipoprotein(a) OC732436|PLGLB2| precursor (Apo(a)) (Lp(a)) | plasminogen-like B2 | TMEM166 transmembrane protein 166 2995476 INMT 1.51E-05 3.47 Indolethylamine N-methyltransferase 3049292 NM_001013398|NM_ IGFBP3 4.56E-05 4.96 Insulin-like growth factor binding protein 3 000598 3058944 NM_001010932|NM_ HGF 1.38E-05 5.69 Hepatocyte growth factor (hepapoietin A; scatter factor) 000601|NM_0010109 31|NM_001010933|N M_001010934 3060332 NM_024636 STEAP4 5.41E-06 3.70 STEAP family member 4 3065740 NM_173054|NM_005 RELN 3.74E-04 5.62 Reelin 045 3095257 NM_194294 INDOL1 2.39E-04 3.08 Indoleamine-pyrrole 2,3 dioxygenase-like 1 3095313 NM_020130 C8orf4 1.82E-07 3.76 Chromosome 8 open reading frame 4 3113133 NM_006438 COLEC10 1.37E-06 13.60 Collectin sub-family member 10 (C-type lectin) 3128817 NM_000680|NM_033 ADRA1A 1.87E-04 3.71 Adrenergic, alpha-1A-, receptor 303|NM_033304|NM_ 033302 3193631 NM_004108|NM_015 FCN2 1.70E-07 3.64 Ficolin (collagen/fibrinogen domain containing lectin) 2 837 (hucolin) 3204285 NM_006274 CCL19 4.46E-05 11.65 Chemokine (C-C motif) ligand 19 3204301 NM_002989 CCL21 1.51E-04 4.04 Chemokine (C-C motif) ligand 21 3215570 NM_000507 FBP1 2.86E-04 3.65 Fructose-1,6-bisphosphatase 1 3237286 NM_001009567 MRC1 1.08E-06 4.72 Mannose receptor, C type 1 3251566 NM_152635 OIT3|C1orf146 2.82E-08 10.95 Oncoprotein induced transcript 3 | chromosome 1 open reading frame 146 3286602 NM_000609|NM_199 CXCL12 3.57E-06 5.02 Chemokine (C-X-C motif) ligand 12 (stromal cell-derived 168|NM_001033886 factor 1) 3302533 NM_003015 SFRP5 1.38E-05 4.31 Secreted frizzled-related protein 5 3333811 NM_001039752 SLC22A10 3.50E-04 5.11 Solute carrier family 22 (organic anion/cation transporter), member 10 3348940 NM_031938|NM_001 KCTD9|BCDO2|LO 3.85E-05 4.40 Potassium channel tetramerisation domain containing 9 | 037290 C728291|LOC64251 beta-carotene dioxygenase 2 | similar to potassium 3|LOC728707|LOC7 channel tetramerisation domain containing 9 | 29404|LOC730942|L hypothetical protein LOC728707 OC650978|LOC6478 52|LOC731540 3349858 NM_006169 NNMT 2.28E-05 9.79 Nicotinamide N-methyltransferase 3359121 NR_003512|NM_0010 INS-IGF2 7.64E-04 13.10 Insulin- insulin-like growth factor 2 | insulin-like growth 07139|NM_000612 factor 2 (somatomedin A) 3360401 NM_000518 HBB 3.13E-06 3.35 Hemoglobin, beta 3362826 NM_006691 XLKD1 6.72E-09 8.65 Extracellular link domain containing 1 3365318 NM_006512 SAA4 9.90E-04 3.56 Serum amyloid A4, constitutive 3374934 NM_152852|NM_022 MS4A6A|LOC64368 2.47E-05 3.25 Membrane-spanning 4-domains, subfamily A, member 349|NM_152851 0 6A | hypothetical protein 3402786 NM_000616 CD4|S100PBP 1.43E-06 3.07
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  • The Immunomodulatory CEA Cell Adhesion Molecule 6 (CEACAM6/Cd66c) Is a Candidate Receptor for the Influenza a Virus

    The Immunomodulatory CEA Cell Adhesion Molecule 6 (CEACAM6/Cd66c) Is a Candidate Receptor for the Influenza a Virus

    bioRxiv preprint doi: https://doi.org/10.1101/104026; this version posted January 30, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 The immunomodulatory CEA cell adhesion molecule 6 (CEACAM6/CD66c) is a 2 candidate receptor for the influenza A virus 3 Shah Kamranur Rahmana *, Mairaj Ahmed Ansarib, Pratibha Gaurc, Imtiyaz Ahmada, 4 Chandrani Chakravartya,d, Dileep Kumar Vermaa, Sanjay Chhibbere, Naila Nehalf, 5 Shanmugaapriya Sellathanbyd, Dagmar Wirthc, Gulam Warisb and Sunil K. Lala,g # 6 7 Virology Group, International Centre for Genetic Engineering & Biotechnology, New Delhi, 8 Indiaa. 9 Department of Microbiology and Immunology, H. M. Bligh Cancer Research Laboratories, 10 Rosalind Franklin University of Medicine and Science, Chicago Medical School, North 11 Chicago, Illinois, USAb. 12 Helmholtz Centre for Infection Research, Braunschweig, Germanyc. 13 Department of Biomedical Science, Bharathidasan University, Trichy, Indiad. 14 Microbiology Department, Panjab University, Chandigarh, Indiae. 15 Career Institute of Medical & Dental Sciences and Hospital, Lucknow, Indiaf. 16 School of Science, Monash University, Selangor DE, Malaysiag. 17 18 Running Head: Protein receptor for Influenza A Virus 19 20 # Corresponding author: Professor of Microbiology, School of Science, Monash University, 21 47500 Bandar Sunway, Selangor DE, Malaysia. 22 Email: [email protected]; Telephone: (+603) 551 59606 23 24 * Current address: Department of Pathogen Molecular Biology, London School of Hygiene & 25 Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom. 26 1 bioRxiv preprint doi: https://doi.org/10.1101/104026; this version posted January 30, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder.
  • MALE Protein Name Accession Number Molecular Weight CP1 CP2 H1 H2 PDAC1 PDAC2 CP Mean H Mean PDAC Mean T-Test PDAC Vs. H T-Test

    MALE Protein Name Accession Number Molecular Weight CP1 CP2 H1 H2 PDAC1 PDAC2 CP Mean H Mean PDAC Mean T-Test PDAC Vs. H T-Test

    MALE t-test t-test Accession Molecular H PDAC PDAC vs. PDAC vs. Protein Name Number Weight CP1 CP2 H1 H2 PDAC1 PDAC2 CP Mean Mean Mean H CP PDAC/H PDAC/CP - 22 kDa protein IPI00219910 22 kDa 7 5 4 8 1 0 6 6 1 0.1126 0.0456 0.1 0.1 - Cold agglutinin FS-1 L-chain (Fragment) IPI00827773 12 kDa 32 39 34 26 53 57 36 30 55 0.0309 0.0388 1.8 1.5 - HRV Fab 027-VL (Fragment) IPI00827643 12 kDa 4 6 0 0 0 0 5 0 0 - 0.0574 - 0.0 - REV25-2 (Fragment) IPI00816794 15 kDa 8 12 5 7 8 9 10 6 8 0.2225 0.3844 1.3 0.8 A1BG Alpha-1B-glycoprotein precursor IPI00022895 54 kDa 115 109 106 112 111 100 112 109 105 0.6497 0.4138 1.0 0.9 A2M Alpha-2-macroglobulin precursor IPI00478003 163 kDa 62 63 86 72 14 18 63 79 16 0.0120 0.0019 0.2 0.3 ABCB1 Multidrug resistance protein 1 IPI00027481 141 kDa 41 46 23 26 52 64 43 25 58 0.0355 0.1660 2.4 1.3 ABHD14B Isoform 1 of Abhydrolase domain-containing proteinIPI00063827 14B 22 kDa 19 15 19 17 15 9 17 18 12 0.2502 0.3306 0.7 0.7 ABP1 Isoform 1 of Amiloride-sensitive amine oxidase [copper-containing]IPI00020982 precursor85 kDa 1 5 8 8 0 0 3 8 0 0.0001 0.2445 0.0 0.0 ACAN aggrecan isoform 2 precursor IPI00027377 250 kDa 38 30 17 28 34 24 34 22 29 0.4877 0.5109 1.3 0.8 ACE Isoform Somatic-1 of Angiotensin-converting enzyme, somaticIPI00437751 isoform precursor150 kDa 48 34 67 56 28 38 41 61 33 0.0600 0.4301 0.5 0.8 ACE2 Isoform 1 of Angiotensin-converting enzyme 2 precursorIPI00465187 92 kDa 11 16 20 30 4 5 13 25 5 0.0557 0.0847 0.2 0.4 ACO1 Cytoplasmic aconitate hydratase IPI00008485 98 kDa 2 2 0 0 0 0 2 0 0 - 0.0081 - 0.0
  • Supplemental Text and Figures

    Supplemental Text and Figures

    Supplemental Materials and Methods Experimental bone metastasis assay Primary PCa cells were sorted by GFP marker from mTmG+ tumors, and 105 cells in 20μL PBS were injected using Hamilton syringe into the tibia of 6-week old NSG mice. Mice were monitored biweekly for moribund signs for euthanasia and organ harvest. Noninvasive mouse and ex vivo imaging MRI imaging with Bruker ICON and fluorescence imaging of fresh organs with metastasis enumeration were recently described (Lu et al. 2017). Primary prostate cell sphere formation assay Isolate of primary cells from prostate, culture and counting of prostatospheres on Matrigel were performed as described (Lukacs et al. 2010). For organoid culture assay, we followed a published matrigel embedding method (Chua et al. 2014). RNA-Seq and differential gene expression Total RNA was isolated from prostate tumors using Direct-zol RNA MiniPrep Kit (Zymo Research) and processed for stranded total RNA-Seq using Illumina HiSeq 4000 at Sequencing and Microarray Facility at MD Anderson Cancer Center. The differential expression analysis was performed using the DESeq2 package of R. P-values obtained after multiple binomial tests were adjusted using BH method. Significant genes are defined by using a cut-off of 0.05 on the BH corrected p-value and an absolute log2 fold change value of at least 1.5. Histology and western blot H&E stain, immunohistochemical (IHC) and western blot were performed as previously described (Ding et al. 2011; Wang et al. 2016). Primary antibodies for IHC include Ki67 (Fisher, RM-9106-S1), cleaved caspase 3 (Cell Signaling Technology aka CST, 9661), cyclin D1 (Fisher, clone SP4), TGFBR2 (Abcam, ab61213), BMPR2 (Abcam, ab130206), AR (EMD Millipore, 06-680), phospho- Akt (CST, 4060), GFP (CST, 2956), E-Cadherin (CST, 14472).