Supplementary Data- Figure legends

S1: Transfection of SH-SY5Y neuroblastoma cells with LSD1-directed siRNA resulted in reproducible knock-down of LSD1 mRNA expression. mRNA levels were measured by quantitative real-time PCR.

S2: TaqMan quantitative RT-PCR confirmed the expression changes detected via microarray analysis for LSD1, DNAL4, DNM2, TNS1 and TPM1. * p < 0.1,** p < 0.5

S3: As TNS1 is induced upon siRNA mediated LSD1 knock-down in vitro, we analyzed TNS1 expression in vivo. TNS1 was significantly down-regulated in primary tumors with low LSD1 expression, and in ganglioneuroma and ganglioneuroblastoma.

S4: Treatment of NB69, SKNAS and Kelly neuroblastoma cells with tranylcypromine, resulted in extensive reduction of cell numbers. Western blot analysis confirmed an accumulation of H3K4 dimethylation upon treatment with tranylcypromine. β-actin served as the loading control.

Supplementary Table 1: Genes annotated with ‚neuronal differentiation’ were analyzed for their correlation with LSD1 expression and differential expression between poorly differentiated neuroblastoma, differentiating neuroblastoma and ganglioneuroma / ganglioneuroblastoma by ANOVA in 110 primary neuroblastic tumors using Affymatrix microarrays. Most genes that showed differential expression between NBpd, NBdiff and GNB/GN were also correlated with LSD1 expression.

Supplementary Table 2: Genes correlating with LSD1 were annotated according to gene ontology. These genes were compared to all genes which have been considered expressed in at least 1 of the 110 samples (the reference) to determine if any GO class was enriched in the genes which correlated with LSD1. P-Values were calculated to evaluate if any go class was significantly overrepresented, and the 5 most significant classes were displayed in table 2.

Supplementary Table 3: Patients were categorized according to known molecular and clinical risk factors. ANOVA was used to analyse if LSD1 is differentially expressed between patients of different groups. While differential expression of LSD1 did not reached significance between patients grouped according to stage, NMYC or

LOH1p36, a week difference was observed between patients grouped according to age.

Supplementary Table 4

Annotation of genes differentially expressed between cells treated with siRNA against

LSD1 and control siRNA. (Probe-IDs correspond to Affymetrix U133v2 microarrays, a=mean expression in control cells, b=mean expression upon siRNA mediated knock down, FC=Fold change, tt=P-Value T-test)

Supplementary data S2: Materials and Methods

LSD1, DNAL4, DNM2, TNS1, TPM1 expression was quantitatively measured using the ABI 7900HT TaqMan instrument (Applied Biosystems, Foster City, CA). TaqMan reactions were done in 384-well plates according to the instructions of the manufacturer. Expression of LSD1, DNAL4, DNM2, TNS1, TPM were measured in duplicates and normalized to 18S RNA. Primers were labelled with 5'-FAM as a reporter and 3'-NFQ-1 as a quencher and purchased from Applied Biosystems.