Supplemental Protocols Protocol 1: RNA isolation Blood collection TM Blood was drawn in PAXgene Blood RNA Tubes, that contain a proprietary reagent composition based on a patented RNA stabilization technology that time stabilizes intracellular RNA. By that, ex vivo changes of expression profiles are avoided. The RNA can be stabilized by PAXgene™ system for three days at 18-25°C, for five days at 2-8°C, and over a longer period (at least 6 months) at -20°C to -70°C. The PAXgene™ tubes contain 6,9 ml of RNA stabilizing solution and are suitable for the sampling of 2,5 ml blood per tube. 2,5 ml to 5 ml blood were sampled from each patient and healthy blood donor. After blood withdrawal either RNA isolation was performed directly or tubes were stored over night at 4°C. For a longer storage over several days tubes were frozen at -20°C. Before RNA isolation the blood was incubated for at least 2 hours at room temperature (RT). Isolation of total RNA from blood cells Blood cells were pelleted by centrifugation (5000g, 10 min, RT). The pellet was resuspended in 10 ml RNase free water, and centrifuged again. The total RNA including the miRNA was isolated and purified using the miRNeasy Mini Kit (Qiagen). The pellet was resuspended in 700 µl QIAzol lysis reagent and incubated for 5 min at RT. Then 140 µl chloroform were added, vortexed for 15 sec, and incubated for 2-3 min at RT. After centrifugation at 14000 rpm and 4°C for 15 min. the upper, watery phase was removed and mixed with 1,5 vol 100% ethanol to precipitate the RNA. Each 700 µl of this mixture were placed on a column and centrifuged at 13000 rpm at RT for 15 sec. After the mixture had completely passed the column, 700 µl of Buffer RWT were added, and centrifuged at 13000 rpm at RT for 15 sec. 500 µl Buffer RPE were added to the column and centrifuged at 13000 rpm at RT for 15 sec. After this, another 500 µl Buffer RPE were added to the column and centrifuged at 13000 rpm at RT for 2 min. The column was then dried by centrifugation at 13000 rpm and RT for 1 min. RNA was eluted with 40 µl RNase free water. The RNA was stored at 70°C until use. Protocol 2: Microarray analysis 300 ng of total RNA was mixed and 1 µl of 5 pM miRNA spike-in mix and dried in a table top speedvac. Each RNA pellet was fully resuspended in 25 µl of this hybridization buffer and denatured for 3 minutes at 95 °C. Until the hybridization, the denatured samples were kept on ice. Microarray hybridization was performed using the Geniom® RT Analyzer and Geniom® miRNA biochips homo sapiens. Samples were loaded automatically and the hybridization (14 hours, 42°C) was started. After hybridization, SAPE solution, antibody solution, equilibration buffer (1xNEB2, New England Biolabs), stop buffer (6xSSPE) and enzyme solution were place into the RT Analyzer. The array equilibration was followed by incubation with enzyme solution. Enzyme incubation was stopped with stop buffer. SAPE staining, signal amplification and detection proceeded fully automated within the Geniom® RT Analyzer. Spike-in mix: 107:5’-GCAAAGGCUAUCGUCAAGAGAUC-3’ 142:5’-GUCGGCAUUUGGCUGGAACUUCAUA-3’ 170:5’-UGACGGGUCUCUUCUUCGAUAGC-3’ 30:5’-CAAAUCAACAAGAUGAGGUCUGGGG-3’ 327:5’-CUUCCUGACCUUACCGAUUCCGA-3’ 35:5’-UCAUUGCCUACAAGCCACCAAGC-3’ 41:5’-GACAAAUCGGAUUCAAGGGCAGG-3’ 46:5’-AGAUGUGGUUGCAACUUCGGAGC-3’ 473:5’-UACCAACCCCACCAAAACCAAACGU-3’ 509:5’-UCCAAAACCAAACCAAAUCCAAACC-3’ 576:5’-ACAACCACUACUUCCGCCGUCAA-3’ 610:5’-AACUCAAGCCGCCGGAAUCUUCA-3’ 63:5’-AACACCCGUCAAGUCCAGUGCAU-3’ 75:5’-UGCGCGGACUCCAACACUUUGUU-3’, 92:5’-UGAUUGUUGUGACACCGGCACUACU-3’ hybridization buffer for eight arrays: 66µl 20x SSPE 22 µl 100% Formamide 22 µl 1x TE buffer 4.4 µl BSA 50 mg/ml 4.4 µl 0.5 % Tween-20 5.5 µl shortmer control oligo mix 1:30 95.7 µl DEPC water shortmer control oligo mix: F2.1-Cy3:5’-[Cy3]TCACTCATGGTTATGGCAGCACTGC-3’ 80 nM F2.2-Bio 5-[bio]GTAGTTCGCCAGTTAATAGTTTGCG-3’ 12 nM F2.3-Bio 5’-[bio]TCTTACCGCTGTTGAGATCCAGTTC-3’ 4 nM F2.4-Bio 5’-[bio]CCCACTCGTGCACCCAACTGATCTT-3’ 0.4 nM F2.5-Bio 5’-[bio]CCATCCAGTCTATTAATTGTTGCCG-3’ 0.04 nM. SAPE solution: 9 ml 6x SSPE 44 µl SAPE 1 mg/ml; 360 µl BSA 50 mg/ml antibody solution: 1750 µl 2x stain buffer (41.7 ml 12x MES; 92.5 ml 5M NaCl; 25 ml Tween-20; 90.8 ml DEPC water) 140 µl BSA 50 mg/ml 35 µl Goat IgG 21 µl biotinylated Anti-streptavidin antibody 1554 µl DEPC water enzyme solution: 44 µl NEB2, 10x 44 µl Bio-dATP, 40 µM 2.9 µl Klenow exo-, 50000 U/ml 349.1 µl DEPC water