Supplemental Figure 1. Generation of RUNX1-RUNX1T1, -catenin and CD200 over-expressing myeloid progenitors. (A) Schematic diagram of the retroviral expression vectors based on PINCO. (B) Representative dot plots showing the immunophenotypic status following retroviral transduction (day 3). (C) In CD200 transduced cells, CD200 protein expression was analysed using human anti-CD200
(M-OX2 clone) by flow cytometry. Quadrants delimit background fluorescence of control-stained-transduced cells.
Supplemental Figure 2. Gene profiles of control (GFP) and RUNX1-RUNX1T1
(RR) transduced human progenitor cells. MASv5.0 data were normalised per gene and per chip and exported into Genespring v7.3 software. Genes determined to be absent from at least 1 paired sample were removed and an unsupervised hierarchical clustering algorithm was performed on the remaining ~14,000 probe sets. Individual samples are shown in columns and genes in rows. The expression levels for individual genes are indicated by colour, as shown in the scale, with red indicating a high level of expression and green depicting a low level of expression. Samples are colour coded. Human progenitor cells grown in media supplemented with IL-3, SCF, G/GM-
CSF and fractionated into 3 main lineages are represented by G, Granulocytes; M,
Monocytes; E, Erythroid.
Supplemental Figure 3. Effect of IL-17 (B or E or in combination) on RUNX1-
RUNX1T1 cell growth. Relative expansion of control (GFP) or RUNX1-RUNX1T1
(RR) transduced cells cultured in IL-3, SCF, G/GM-CSF and IL-17 (500 ng/ml). Data indicates mean1SD (n=2).
1 Supplemental Table 1. Clinical and biological characteristics of patients and healthy donors tested in this study. AAge at diagnosis and entry into the study.
BFlt3, Fms like tyrosine kinase 3. n/d, not detected.
Supplemental Table 2. Identification of dysregulated probe sets in day 3 and day
6 RUNX1-RUNX1T1 transduced cells. The expressed genes in RUNX1-RUNX1T1 transduced human progenitor cells are listed according to their expression level with respect to control. Individual genes are represented by numbered probes sets on the
Affymetrix HG-U133A oligonucleotide GeneChip. Identification of dysregulated probe sets in CD34+ mixed progenitor cells (n=465), day 6 granulocytes (n=79), day 6 monocytes (n=288) and day 6 erythroid cells (n=209) is described in Methods and
Figure 2. Data on gene function and name were extracted from the gene ontology database via Genespring software.
Supplemental Table 3. Myeloid colony growth of human CD34+ cells expressing
RUNX1-RUNX1T1 or CD200. The data summarises the effect of IL-17B, IL-17E
(or in combination) on colony forming efficiency (as described in Materials and
Methods) in RUNX1-RUNX1T1 transduced cells. Also shown is the effect of over- expressing CD200 on colony forming efficiency. Retrovirally transduced (day 3) cells were grown in IL-3, SCF and G/GM-CSF with or without the addition of 1, 50 or 500 ng/ml IL-17. Colony formation was scored after 7 Days. Data indicates mean percentage values ± SD (n=3). Nd; not done.
2 Supplemental Table 4. Identification of 38 dysregulated probe sets common to day 3 and day 6 RUNX1-RUNX1T1 transduced monocytes. The expressed genes in RUNX1-RUNX1T1 transduced human progenitor cells are listed according to their expression level with respect to the control on day 6. Individual genes are represented by numbered probes sets on the Affymetrix HG-U133A oligonucleotide GeneChip.
Over lap of dysregulated genes in other lineages (Table 1 and Supplemental Table 5) are denoted by G, granulocytes; E, erythroid and M, mononcytes. AInconsistent gene regulation when compared to day 3 data set (list #1). Data on gene function and name were extracted from the gene ontology database via Genespring software.
Supplemental Table 5. Identification of 64 dysregulated probe sets common to day 3 and day 6 RUNX1-RUNX1T1 transduced erythroid cells. The expressed genes in RUNX1-RUNX1T1 transduced human progenitor cells are listed according to their expression level with respect to control on day 6. Individual genes are represented by numbered probes sets on the Affymetrix HG-U133A oligonucleotide
GeneChip. Over lap of dysregulated genes in other lineages (Table 1 and
Supplemental Table 4) are denoted by G, granulocytes; E, erythroid and M, mononcytes. AInconsistent gene regulation when compared to day 3 data set (list #1).
Data on gene function and name were extracted from the gene ontology database via
Genespring software.
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