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Fig. S1. Detection of EWSR1–ETS fusion transcripts in Ewing family tumor (EFT) samples. Presented are representative results from RT-PCR analysis of EFT cell lines (fromleft to right: SK-N-MC, A673, RD-ES, SK-ES, STA-ET8, TC-32, TC-71, TTC-466). β-Actin (ACTB) was used as a control. NTC, no template control. With the exception ofone cell line that was positive for EWS–ERG, all cell lines expressedEWSR1–FLI1 (fusion types I or II).

Fig. S2. AbsenceAUTHOR: Please check that this figureis correct as it looks to be corrupted of aristaless (ARX) expression inEpstein–Barr virus-immortalized B cells. Presented are results fromquantitative RT-PCR analysis with α/β hydrolase domain containing 6 (ABHD6)- and ARX-specific primers. The Ewing family tumor (EFT) cell line SK-N-MC wasused as a positive control. The neuroblastoma cell line SH-SY5Y was used as anegative control and set as one. Squares and triangles represent conditionallyEpstein–Barr virus-immortalized B cell line P493-6. Squares represent cellscultured in the presence of estrogen (+E). Triangles represent cells culturedin the absence of estrogen (w/o E). Open squares and triangles represent P493-6cells cultured in the presence of tetracycline (MYC off), closed squares andtriangles represent P493-6 cells cultured in the absence of tetracycline (MYCon).

Fig. S3. Phylogenetic analysis of α/β hydrolasedomain containing 6 (ABHD6) in chordata. (A) Neighbor joiningtree of protein sequences of ABHD6 in different species. (B) Consensusmotif of ABHD6 homologues. Sequence logos were generated on the basis ofABHD6 sequences from the species shown in (A). All of these enzymesencode the central serine by the codon TCN.

Fig. S4. Alignment of α/β hydrolase domain containing6 (ABHD6) protein sequences from different chordate species. Presented aresequences of the catalytic core of ABHD6 in the indicated species (from Tyr58to Phe323 of the human ABHD6 sequence). Alignment was calculated with ClustelWand visualized with GeneDoc (Nicholas, K. B., and Nicholas H.B. Jr. 1997. GeneDoc: a tool for editing and annotation multiple sequence alignements. Distributedby the author).

Fig. S5. Sequence logo of α/β hydrolase domaincontaining 6 (ABHD6). The sequence logo was created on the basis of thesequences from Supplementary Fig.  S4. The amino acids of thecatalytic triad are indicated by boxes.

Fig. S6. Predicted structure of human α/β hydrolasedomain containing 6 (ABHD6). The structure of human ABHD6 (green) was predictedby homology modeling using the α/β hydrolase homoserine transacetylase of H. influenzae (white) as model. The aminoacids of the catalytic triad are indicated. Both structures are characterizedby the presence of two domains. The contact zone between both domains forms a“catalytic tunnel”. See Discussion in the text.

Fig. S7. Expression of α/β hydrolase domaincontaining 6 (ABHD6) in Ewing family tumor (EFT) samples did not correlate withclinical prognosis. Presented are signal intensities from two data sets (HG_U133A microarrays: Schaefer KL, Eisenacher M, Braun Y et  al.Eur J Cancer. 2008; 44: 699-709; HG_U133Plus2.0 microarrays: Scotlandi K, Remondini D, Castellani G et  al. J Clin Oncol 2009; 27: 2209–16) including patients with (closed bars) and without (open bars) relapses. Bars at the right represent means and standard deviations.

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