Targets of SOX17 detected by ChIP-on-chip analysis. (A) GO analysis of the Sox17 targets detected by ChIP-on-chip analysis. CD34⁺CD43⁻ cells from day 6 EBs were transduced with a 3× Flag SOX17-ERT retrovirus. The cells were further cultured on OP9 cells in the presence of 20 ng/mL of SCF and TPO and 200nM 4-OHT. CD34⁺ cells were collected at day 27 of culture and subjected to ChIP-on-chip analysis. P for each GO term is indicated. (B) ChIP-on-chip profile of SOX17 occupancy at genes related to hematopoietic development from HE cells. Plot under the x-axis shows the position of probe sets. Arrowhead at the VE-cadherin/CDH5 promoter indicate consensus motif of Sox17-binding site. (C) Gene-expression patterns of wild-type and engineered cells obtained in microarray analyses clustered using hierarchical clustering. Distance between 2 samples was defined with the Pearson correlation of Sox17 target genes with Sox17 binding more than 3-fold in the ChIP-on-chip analysis presented in supplemental Table 2. The color of each cell represents the value of correlation indicated on the right side of the matrix. (D) Comparative analysis of ChIP-on-chip and microarray data. Venn diagrams showing the number of genes bound by Sox17 (> 3-fold enrichment) and the number of genes up-regulated in expression more than 2-fold in at least 1 cell type among ECs, pre-HPCs, and HPCs on overexpression of Sox17 (Sox17-overexpressing CD34⁺CD43⁺CD45^−/low cells compared with those of respective fresh controls; left panel). The percentages of overlapping and nonoverlapping bound genes are indicated in parentheses. Shown are the correlation of Sox17 binding (fold enrichment) in ChIP-on-chip analysis and the fold changes in expression during differentiation of ECs to HPCs (right panel). The fold enrichments and fold changes in expression were plotted for 84 genes of 98 showing enrichments greater than 3-fold (the microarray data were not available for the remaining 14 genes). Correlation coefficients (R) are indicated for genes with fold enrichment greater than 3- and 6-fold, respectively.