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1 Fujisaki Et Al. EXPANSION of HIGHLY CYTOTOXIC HUMAN

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1 Fujisaki Et Al. EXPANSION of HIGHLY CYTOTOXIC HUMAN Fujisaki et al. EXPANSION OF HIGHLY CYTOTOXIC HUMAN NATURAL KILLER CELLS FOR CANCER CELL THERAPY Supplementary Tables 1-6 Supplementary Figures 1-5 1 Supplementary Table 1. List of antibodies used in this study Antigen Fluorochromea Sourceb CD56 FITC BD APC Beckman Coulter CD3 PerCP BD ECD Beckman Coulter KIR2DL1 (CD158a) PE BD and Beckman Coulter KIR2DL2/3 (CD158b) FITC BD PE BD KIR2DL3 (NKAT2) FITC BD KIR3DL1 (CD158e, NKB1) PE BD and Miltenyi Biotec NKp30 (CD337) PE Beckman Coulter NKp44 (CD336) PE Beckman Coulter NKp46 (CD335) PE Beckman Coulter NKG2D (CD314) PE Beckman Coulter MIC A/B PE BD ULBP-1 Unconjugated R&D Systems ULBP-2 PE R&D Systems ULBP-3 Unconjugated R&D Systems IL-2RD (CD25) PE Dako CXCR6 (CD186) PE R&D Systems TNFSF4 (CD252, OX40L) PE BD TNFSF7 (CD70) PE BD ITGA2 (CD49b) PE BD ITGB7 PE BD VCAM1 (CD106) PE BD Mouse IgG1 PE BD Mouse IgG2a PE BD Mouse IgG2b PE BD Mouse IgG3 PE BD aFITC, fluorescein isothiocyanate; APC, allophycocyanin; PerCP, peridin chrophyll protein; ECD, phycoerythrin-Texas Red; PE, phycoerythrin. Unconjugated antibodies were visualized with a goat- anti-mouse IgG2a PE (Southern Biotechnology Assoc., Birmingham, AL). bBD (Becton Dickinson, San Jose, CA); Beckman Coulter, Miami, FL; Miltenyi Biotec, Auburn, CA; R&D Systems, Minneapolis, MN; Dako, Carpinteria, CA 2 Supplementary Table 2. Top 100 probesets overexpressed in NKAES NK cells (n = 5) as compared to unstimulated NK cells (n = 5) ranked by fold expression ProbeSet Gene Gene Title FDR P-Value Fold* Symbol 202095_s_at BIRC5 baculoviral IAP repeat-containing 5 (survivin) 0.0000 0.0000 229.2 203716_s_at DPP4 dipeptidyl-peptidase 4 (CD26, adenosine 0.0001 0.0000 171.2 deaminase complexing protein 2) 222958_s_at DEPDC1 DEP domain containing 1 0.0000 0.0000 155.5 228868_x_at CDT1 Chromatin licensing and DNA replication factor 1 0.0001 0.0000 140.4 218755_at KIF20A kinesin family member 20A 0.0005 0.0000 128.9 232278_s_at DEPDC1 DEP domain containing 1 0.0004 0.0000 127.8 232596_at DIAPH3 diaphanous homolog 3 (Drosophila) 0.0000 0.0000 125.8 215509_s_at BUB1 BUB1 budding uninhibited by benzimidazoles 1 0.0004 0.0000 115.4 homolog (yeast) 202705_at CCNB2 cyclin B2 0.0000 0.0000 110.7 205164_at GCAT glycine C-acetyltransferase (2-amino-3- 0.0000 0.0000 110.4 ketobutyrate coenzyme A ligase) 212949_at NCAPH non-SMC condensin I complex, subunit H 0.0001 0.0000 109.7 204126_s_at CDC45L CDC45 cell division cycle 45-like (S. cerevisiae) 0.0002 0.0000 108.2 202870_s_at CDC20 cell division cycle 20 homolog (S. cerevisiae) 0.0000 0.0000 107.1 210229_s_at CSF2 colony stimulating factor 2 (granulocyte- 0.0006 0.0000 101.6 macrophage) 209891_at SPC25 SPC25, NDC80 kinetochore complex component, 0.0001 0.0000 100.5 homolog (S. cerevisiae) 203868_s_at VCAM1 vascular cell adhesion molecule 1 0.0014 0.0001 93.7 209369_at ANXA3 annexin A3 0.0008 0.0000 87.3 203418_at CCNA2 cyclin A2 0.0001 0.0000 86.8 232397_at --- Homo sapiens, clone IMAGE;4332461, mRNA 0.0008 0.0000 86.7 219148_at PBK PDZ binding kinase 0.0001 0.0000 85.0 204695_at CDC25A cell division cycle 25 homolog A (S. pombe) 0.0000 0.0000 84.3 218542_at CEP55 centrosomal protein 55kDa 0.0000 0.0000 83.0 212022_s_at MKI67 antigen identified by monoclonal antibody Ki-67 0.0000 0.0000 82.9 220651_s_at MCM10 minichromosome maintenance complex 0.0006 0.0000 82.5 component 10 209680_s_at KIFC1 kinesin family member C1 0.0000 0.0000 80.5 211080_s_at NEK2 NIMA (never in mitosis gene a)-related kinase 2 0.0001 0.0000 80.0 203213_at CDC2 cell division cycle 2, G1 to S and G2 to M 0.0000 0.0000 79.4 219990_at E2F8 E2F transcription factor 8 0.0001 0.0000 77.3 1554408_a_at TK1 thymidine kinase 1, soluble 0.0001 0.0000 76.2 209773_s_at RRM2 ribonucleotide reductase M2 polypeptide 0.0000 0.0000 75.4 209832_s_at CDT1 chromatin licensing and DNA replication factor 1 0.0002 0.0000 73.7 204641_at NEK2 NIMA (never in mitosis gene a)-related kinase 2 0.0003 0.0000 73.5 201292_at TOP2A topoisomerase (DNA) II alpha 170kDa 0.0000 0.0000 71.6 203764_at DLG7 discs, large homolog 7 (Drosophila) 0.0000 0.0000 71.3 202338_at TK1 thymidine kinase 1, soluble 0.0000 0.0000 71.3 211478_s_at DPP4 dipeptidyl-peptidase 4 (CD26, adenosine 0.0005 0.0000 66.8 deaminase complexing protein 2) 219918_s_at ASPM asp (abnormal spindle) homolog, microcephaly 0.0000 0.0000 66.6 associated (Drosophila) 3 221795_at NTRK2 neurotrophic tyrosine kinase, receptor, type 2 0.0001 0.0000 63.5 203214_x_at CDC2 cell division cycle 2, G1 to S and G2 to M 0.0000 0.0000 61.9 1555758_a_at CDKN3 cyclin-dependent kinase inhibitor 3 (CDK2- 0.0000 0.0000 60.4 associated dual specificity phosphatase) 201890_at RRM2 ribonucleotide reductase M2 polypeptide 0.0000 0.0000 59.6 210559_s_at CDC2 cell division cycle 2, G1 to S and G2 to M 0.0000 0.0000 59.0 221521_s_at GINS2 GINS complex subunit 2 (Psf2 homolog) 0.0000 0.0000 58.8 204709_s_at KIF23 kinesin family member 23 0.0000 0.0000 58.3 201291_s_at TOP2A topoisomerase (DNA) II alpha 170kDa 0.0000 0.0000 57.9 229610_at CKAP2L cytoskeleton associated protein 2-like 0.0000 0.0000 57.3 203755_at BUB1B BUB1 budding uninhibited by benzimidazoles 1 0.0001 0.0000 55.9 homolog beta (yeast) 205024_s_at RAD51 RAD51 homolog (RecA homolog, E. coli) (S. 0.0001 0.0000 55.8 cerevisiae) 209464_at AURKB aurora kinase B 0.0000 0.0000 55.5 204825_at MELK maternal embryonic leucine zipper kinase 0.0000 0.0000 55.1 204962_s_at CENPA centromere protein A 0.0000 0.0000 54.0 222962_s_at MCM10 minichromosome maintenance complex 0.0004 0.0000 53.1 component 10 220295_x_at DEPDC1; DEP domain containing 1; DEP domain containing 0.0000 0.0000 53.0 LOC7308 1 pseudogene 88 1554696_s_at TYMS thymidylate synthetase 0.0000 0.0000 52.4 207165_at HMMR hyaluronan-mediated motility receptor (RHAMM) 0.0000 0.0000 51.6 1556046_a_at LOC1576 hypothetical protein LOC157627 0.0001 0.0000 51.5 27 202589_at TYMS thymidylate synthetase 0.0000 0.0000 51.1 236641_at KIF14 kinesin family member 14 0.0000 0.0000 48.9 206364_at KIF14 kinesin family member 14 0.0000 0.0000 48.8 218585_s_at DTL denticleless homolog (Drosophila) 0.0000 0.0000 47.7 201710_at MYBL2 v-myb myeloblastosis viral oncogene homolog 0.0000 0.0000 47.4 (avian)-like 2 225655_at UHRF1 ubiquitin-like, containing PHD and RING finger 0.0000 0.0000 46.7 domains, 1 219493_at SHCBP1 SHC SH2-domain binding protein 1 0.0000 0.0000 46.5 222608_s_at ANLN anillin, actin binding protein 0.0000 0.0000 45.7 229490_s_at --- Transcribed locus 0.0007 0.0000 45.2 218663_at NCAPG non-SMC condensin I complex, subunit G 0.0000 0.0000 45.1 202240_at PLK1 polo-like kinase 1 (Drosophila) 0.0000 0.0000 44.9 208343_s_at NR5A2 nuclear receptor subfamily 5, group A, member 2 0.0008 0.0000 44.5 209709_s_at HMMR hyaluronan-mediated motility receptor (RHAMM) 0.0000 0.0000 44.4 210052_s_at TPX2 TPX2, microtubule-associated, homolog (Xenopus 0.0000 0.0000 44.3 laevis) 203717_at DPP4 dipeptidyl-peptidase 4 (CD26, adenosine 0.0002 0.0000 44.1 deaminase complexing protein 2) 212023_s_at MKI67 antigen identified by monoclonal antibody Ki-67 0.0000 0.0000 43.5 218726_at HJURP Holliday junction recognition protein 0.0001 0.0000 43.2 202580_x_at FOXM1 forkhead box M1 0.0001 0.0000 42.9 239219_at AURKB aurora kinase B 0.0002 0.0000 42.8 202954_at UBE2C ubiquitin-conjugating enzyme E2C 0.0000 0.0000 42.4 228729_at CCNB1 cyclin B1 0.0010 0.0001 42.2 4 220085_at HELLS helicase, lymphoid-specific 0.0013 0.0001 41.3 214839_at LOC1576 hypothetical protein LOC157627 0.0001 0.0000 40.6 27 202503_s_at KIAA010 KIAA0101 0.0000 0.0000 40.5 1 209714_s_at CDKN3 cyclin-dependent kinase inhibitor 3 (CDK2- 0.0000 0.0000 40.4 associated dual specificity phosphatase) 238532_at DPF3 D4, zinc and double PHD fingers, family 3 0.0014 0.0001 40.2 218039_at NUSAP1 nucleolar and spindle associated protein 1 0.0000 0.0000 39.9 209642_at BUB1 BUB1 budding uninhibited by benzimidazoles 1 0.0000 0.0000 39.7 homolog (yeast) 214710_s_at CCNB1 cyclin B1 0.0000 0.0000 39.4 213599_at OIP5 Opa interacting protein 5 0.0000 0.0000 39.2 224204_x_at ARNTL2 aryl hydrocarbon receptor nuclear translocator-like 0.0017 0.0001 38.8 2 226829_at AFAP1L2 actin filament associated protein 1-like 2 0.0000 0.0000 38.8 204603_at EXO1 exonuclease 1 0.0001 0.0000 38.7 219000_s_at DCC1 defective in sister chromatid cohesion homolog 1 0.0000 0.0000 38.5 (S. cerevisiae) 219978_s_at NUSAP1 nucleolar and spindle associated protein 1 0.0000 0.0000 38.4 205032_at ITGA2 integrin, alpha 2 (CD49B, alpha 2 subunit of VLA- 0.0020 0.0002 37.7 2 receptor) 223062_s_at PSAT1 phosphoserine aminotransferase 1 0.0000 0.0000 37.6 207533_at CCL1 chemokine (C-C motif) ligand 1 0.0034 0.0004 37.4 223700_at MND1 meiotic nuclear divisions 1 homolog (S. 0.0001 0.0000 37.4 cerevisiae) 219650_at ERCC6L excision repair cross-complementing rodent repair 0.0006 0.0000 36.9 deficiency, complementation group 6-like 218662_s_at NCAPG non-SMC condensin I complex, subunit G 0.0000 0.0000 36.7 204558_at RAD54L RAD54-like (S. cerevisiae) 0.0000 0.0000 36.4 209408_at KIF2C kinesin family member 2C 0.0000 0.0000 35.5 210334_x_at BIRC5 baculoviral IAP repeat-containing 5 (survivin) 0.0010 0.0001 34.9 * Fold expression in NKAES-NK cells compared to unstimulated NK cells.
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