Table 1: Differentially Expressed Proteins in BM and PB Plasma Derived from Patients At

Table 1: Differentially expressed proteins in BM and PB plasma derived from patients at MDS, AML diagnosis and three months following treatment (FT). Sample Protein Protein entry Name/Identity MW PI Mascot Matched Coverage Spot No. Expression identity Acces. Score Peptides % %total density level No. (x1000) Patients Controls BMP/MDS P02774 VTDB_HUMAN 54525.00 5.28 154 20/92 48 23±1.1 41±2.9 0.41* Vitamin D binding protein precursor BMP/MDS P01008 ANT3_HUMAN 53025.00 6.30 110 19/76 42 27±2.1 65±1.3 0.63* Antithrombin III precursor BMP/MDS P00450 CERU_HUMAN 122983.00 5.40 161 26/82 29 13±1.9 31±2.7 0.64* Ceruloplasmin precursor BMP/MDS P00739 HPTR_HUMAN 39496.00 6.46 92 14/82 36 51±2.3 27±3.1 1.71** Haptoglobin related protein precursor BMP/AML• P10909 CLUS_HUMAN 53031.00 5.90 82 12/54 32 47±2.5 29±3.1 1.90** Clusterin precursor BMP/AML P25311 ZA2G_HUMAN 34079.00 4.50 114 20/89 58 28±1.6 14±1.5 1.83** Zinc-alpha-2 glycoprotein BMP/AML P02743 SAMP_HUMAN 25485.00 6.10 69 8/57 30 44±1.7 25±2.5 1.35** Serum amyloid P component precursor BMP/AML P02735 SAA_HUMAN 13581.00 6.40 94 9/83 70 47±2.4 26±2.8 1.28** Serum amyloid A protein precursor BMP/AML P02647 APOA1_HUMAN 30759.00 5.50 102 17/72 50 14±1.8 33±1.9 0.71** Apolipoprotein A1 precursor BMP/AML P02655 APOC2_HUMAN 11257.00 4.50 59 6/49 52 17±2.8 39±2.1 0.77** Apolipoprotein C-II precursor BMP/AML P01011 AACT_HUMAN 46878.00 5.30 212 26/92 53 11±2.5 37±1.9 0.64** Alpha-1 antichymotrypsin precursor BMP/AML P04217 A1BG_HUMAN 54809.00 5.50 149 18/78 41 59±2.1 42±1.9 1.98* Alpha-1B glycoprotein precursor BMP/AML P02765 FETUA_HUMAN 400098.00 5.36 57 8/55 21 15±2.3 36±1.7 0.28** Alpha-2-HS glycoprotein precursor BMP/AML P02760 AMBP_HUMAN 39886.00 5.90 76 14/83 34 56±3.1 38±2 1.21* AMBP protein precursor BMP/AML P00738 HPT_HUMAN 45861.00 6.10 140 19/82 38 49±2.9 27±2.4 2.09** Haptoglobin precursor BMP/AML P69905 HBA_HUMAN 16102.00 6.90 183 12/70 88 40±1.1 27±2.8 1.14** Hemoglobin subunit alpha BMP/AML O43866 CD5L_HUMAN 39603.00 5.20 125 16/88 54 28±2.1 48±2.8 0.63** CD5 antigen- like precursor BMP/AML P02750 A2GL_HUMAN 38382.00 6.50 65 9/74 41 24±2.3 51±1.4 0.90** Leucine rich alpha 2 glycoprotein precursor BMP/AML P09871 C1S_HUMAN 78174.00 4.70 168 23/63 36 12±1.9 36±2.4 0.48** Complement C1s subcomponent precursor BMP/AML P00736 C1R_HUMAN 81661.00 5.86 112 21/84 39 49±1.8 26±2.7 1.33** Complement C1r subcomponent prcursor BMP/AML P00751 CFAB_HUMAN 86847.00 6.70 128 18/88 35 35±1.9 11±3.4 1.29** Complement factor B BMP/FT P02774 VTDB_HUMAN 54525.00 5.32 190 26/91 58 39±3.7 18±2.5 1.68*

The identified proteins are designated with their abbreviated names and the Swiss-Prot accession numbers. The theoretical molecular weight (MW) calculated by CalPI/MW; available in the Swiss-Prot Web site, the theoretical isoelectric point (pI), the probability of a random identification (Mascot score), the number of the peptides used for identification and coverage of each protein are also demonstrated. Score is 10*log (p), where P the probability of the observed match to be a random event. Mascot scores >55 indicate identity, or extensive homology at the p<0.05 level. Expression levels >1 suggest overexpression, whilst those <1 indicate suppression (*p, 0.05, **p, 0.005).Where: BMP; BM plasma and PBP; PB plasma. •AML; consider both MDS/AML and de novo AML Table 2: Differentially expressed proteins in BM and PB cell lysates derived from patients at MDS and AML diagnoses and three months following treatment (FT). Sample Protein Protein entry Name/Identity MW PI Mascot Matched Coverage Spot No. %total Expression identity Acces. Score Peptides % density (x1000) level No. Patients Controls BMC/MDS P11021 GRP78_HUMAN 72402.00 4.90 89 15/57 24 46±1 21±2.3 1.28* 78 kDa glucose-regulated protein BMC/AML• P25705 ATPA_HUMAN 59828.00 9.60 71 9/33 22 23±2 69±3 0.98** ATP synthase subunit alpha BMC/AML P11021 GRP78_HUMAN 72402.00 4.92 65 8/68 18 52±1 16±2.7 1.65** 78 kDa glucose-regulated protein BMC/AML P60709 ACTB_HUMAN 42052.00 5.20 93 9/68 36 15±1 39±2.1 0.36* Actin cytoplasmic 1 BMC/AML P63061 ACTG_HUMAN 42108.00 5.20 93 9/68 37 19±3 33±2 0.62** Actin cytoplasmic 2 BMC/AML O95831 AIFM1_HUMAN 67144.00 9.60 68 7/22 15 39±2.1 12±1 1.74* Apoptosis inducing factor 1 mitochondrial BMC/AML P13645 K1C10-HUMAN 59020.00 5.00 57 5/35 13 28±2.5 14±1.6 1.52** Keratin type 1 cytoskeletal 10 BMC/AML P34931 HSP71_HUMAN 70294.00 5.40 65 8/29 16 48±3 17±3.4 1.33* Heat shock 70 kDa protein 1A BMC/AML P09622 DLDH_HUMAN 54713.00 8.90 164 5/18 12 14±2.6 35±1.3 0.85** Dihydrolipoyl dehydrogenase mitochondrial BMC/AML P11142 HSP7C_HUMAN 71082.00 5.20 73 7/36 15 36±1.6 19±2.8 1.69** Heat shock cognate 71 kDa protein BMC/AML P49411 EFTU_HUMAN Elongation factor Tu 49852.00 7.90 165 7/22 16 11±3.2 37±2.4 0.69* mitochondrial BMC/AML P00367 DHE3_HUMAN 61701.00 8.50 130 20/58 33 17±2.9 39±2.3 0.36** Glutamate dehydrogenase 1 mitochondrial BMC/AML P04179 SODM_HUMAN 24878.00 9.10 75 6/21 36 16±2 33±3.6 0.33** Superoxide dismutase [Mn] mitochondrial BMC/AML P02545 LMNA_HUMAN 74380.00 6.60 56 6/20 8 54±2.5 28±3.5 1.21** Lamin A/C BMC/AML P28331 NDUS1_HUMAN 80443.00 5.80 168 8/26 17 61±1.7 38±4 1.54** NADH-ubiquinone oxidoreductase 75kDa subunit BMC/AML P055084 ECHB_HUMAN 51547.00 10.0 57 6/32 15 10±3.1 41±4 0.19** Trifunctional enzyme subunit beta BMC/AML P49448 DHE4_HUMAN 61738.00 9.26 168 12/58 24 57±2 34±2.4 2.03** Glutamate dehydrogenase 2 mitochondrial BMC/FT P27797 CALR_HUMAN 48283.00 4.10 60 6/22 13 12±1.6 45±3 0.29* Calreticulin BMC/FT P13645 K1C10-HUMAN 59020.00 5.00 98 8/61 19 55±2.3 28±3 1.23* Keratin type 1 cytoskeletal 10 PBC/MDS P13645 K1C10-HUMAN 59020.00 5.01 67 6/58 18 28±1 11±2.9 1.29** Keratin type 1 cytoskeletal 10 PBC/AML P06733 ENOA_HUMAN 47481.00 7.70 64 8/29 22 33±2.2 17±3.4 1.45** Alpha enolase PBC/AML P13645 K1C10-HUMAN 59020.00 5.00 77 5/57 16 34±1.7 17±2.9 1.24** The identified proteins are designated with their abbreviated names and the Swiss-Prot accession numbers. The theoretical molecular weight (MW) calculated by CalPI/MW; available in the Swiss-Prot Web site, the theoretical isoelectric point (pI), the probability of a random identification (Mascot score), the number of the peptides used for identification and coverage of each protein are also demonstrated. Score is 10*log (p), where P the probability of the observed match to be a random event. Mascot scores >55 indicate identity, or extensive homology at the p<0.05 level. Expression levels >1 suggest overexpression, whilst those <1 indicate suppression (*p, 0.05, **p, 0.005).Where: BMC; BM cells and PBC; PB cells. •AML; consider both MDS/AML and de novo AML Table 3: Differentially expressed pre-fractionated proteins in BM and PB plasma derived from AML patients at diagnosIs. Sample identity Protein Protein entry Name/Identity MW PI Mascot Matched Coverage Spot No. %total Expression Acces. Score Peptides % density (x1000) level No. Patients Controls PF-BMP/AML• Q03591 FHR1_HUMAN 38766.00 8.70 63 7/53 63 52±2 36±2.1 1.27* Complement factor H related protein 1 PF-BMP/AML Q9UJ70 NAGK_HUMAN 37694.00 5.80 103 9/37 29 62±1 24±3 1.22** N-acetyl-D-glucosamine kinase PF-PBP/AML P06733 ENOA_HUMAN 47481.00 7.70 186 15/49 50 28±2 11±1 1.75* Alpha enolase PF-PBP/AML P10809 CH60_HUMAN 61187.00 5.60 101 10/65 28 36±3 14±2.3 1.14* 60kDa heat shock protein, mitochondrial PF-PBP/AML P34931 HSP71_HUMAN 70294.00 5.40 187 20/82 42 39±2 25±1 1.29* Heat shock 70 kDa protein 1A PF-PBP/AML P11142 HSP7C_HUMAN 71082.00 5.20 178 21/88 41 45±1 12±2.3 1.63* Heat shock cognate 71 kDa protein PF-PBP/AML P04040 CATA_HUMAN 59947.00 7.00 252 23/76 53 62±3 41±1.2 1.26* Catalase PF-PBP/AML P11021 GRP78_HUMAN 72402.00 4.90 282 26/86 52 38±2 30±1 1.18* 78 kDa glucose-regulated protein PF-PBP/AML P30101 PDIA3_HUMAN 57146.00 5.90 166 17/72 40 60±2 40±3.5 1.36* Protein disulfide isomerase A3 PF-PBP/AML Q9UJ72 ANX10_HUMAN 38918.00 6.70 142 12/72 51 39±1 21±0.3 1.52* Annexin A10 PF-PBP/AML P04003 C4BPA_HUMAN 69042.00 7.90 231 22/89 50 23±3 52±2.3 0.63* C4b-binding protein alpha chain PF-PBP/AML Q16555 DPYL2_HUMAN 62711.00 5.90 133 12/44 28 12±1 44±2 0.91* Dihydropyrimidinase-related protein-2 PF-PBP/AML P30040 ERP29_HUMAN 29032.00 7.50 92 8/32 30 18±3 31±1.5 0.25* Endoplasmic reticulum protein ERp29 PF-PBP/AML P15311 EZRI_HUMAN 69484.00 5.40 67 8/47 16 54±2 32±1.7 1.63* Ezrin PF-PBP/AML P52907 CAZA1_HUMAN 33073.00 5.40 108 9/50 44 11±3 38±1 0.38** F-actin-capping protein subunit alpha-1 PF-PBP/AML P47756 CAPZB_HUMAN 31616.00 5.20 63 5/29 22 22±3 70±1.4 0.29* F-actin-capping protein subunit beta

PF-PBP/AML P04406 G3P_HUMAN 36201.00 9.30 87 8/51 35 14±1 29±3.1 0.25** Glyceraldehyde-3 phosphate dehydrogenase PF-PBP/AML P61978 HNRPK_HUMAN 51230.00 5.30 63 7/51 18 19±3 48±1 0.69* Heterogeneous nuclear ribonucleoprotein K PF-PBP/AML P01871 IGHM_HUMAN 49960.00 6.40 93 11/74 28 52±2 15±1.9 1.84** Ig mu chain C region PF-PBP/AML P04220 MUCB_HUMAN 43543.00 4.99 65 21/77 24 62±1 14±2.5 1.59* Ig mu heavy chain disease protein PF-PBP/AML Q14624 ITIH4_HUMAN 103521.00 6.50 166 22/80 26 19±2 54±0.3 0.51* Inter- alpha trypsin inhibitor heavy chain H4 PF-PBP/AML Q06323 PSME1_HUMAN 28876.00 5.70 90 8/51 36 11±1 25±1.9 0.56** Proteasome activator complex subunit 1 The identified proteins are designated with their abbreviated names and the Swiss-Prot accession numbers. The theoretical molecular weight (MW) calculated by CalPI/MW; available in the Swiss-Prot Web site, the theoretical isoelectric point (pI), the probability of a random identification (Mascot score), the number of the peptides used for identification and coverage of each protein are also demonstrated. Score is 10*log (p), where P the probability of the observed match to be a random event. Mascot scores >55 indicate identity, or extensive homology at the p<0.05 level. Expression levels >1 suggest overexpression, whilst those <1 indicate suppression (*p, 0.05, **p, 0.005).Where: PF-BMP; pre-fractionated BM plasma and PF-PBP; pre-fractionated PB plasma. •AML; consider both MDS/AML and de novo AML.

Netwotrk Protein Analysis. Differentially expressed proteins, identified in the present study, were used for pathway analysis. For this purpose, the Swiss-Prot accession numbers were inserted into the String software available at http://string.embl.de * REFERENCE: Jensen LJ, Kuhn M, Stark M, Chaffron S, Creevey C, Muller J, et al.. STRING 8--a global view on proteins and their functional interactions in 630 organisms. Nucleic Acids Res. 2009, 37(Database issue): D412-416. B A

Figure 4: Expression Profiling Diagrams of the BM & PB plasma overexpressed proteins in A. MDS and B. AML, respectively. The diagrams were constructed with the use of the String software. Figure 5: Western blot analysis of MOES (68kDa) expression in de novo AML patients (Lanes 1 and 2), MDS/AML patients (Lanes 3 and 4) and non leukemic patients (Lanes 5-8).