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. 2009 Aug 21;4(8):e6719. doi: 10.1371/journal.pone.0006719

Figure 11. Homeodomain-independent gene regulation by NUP98-HOXA9 in primary human CD34+ cells.

Figure 11

(A) Primary human CD34+ cells were retrovirally transduced with either control MSCV-IRES-GFP vector or vector expressing NUP98-HOXA9 (NH9), NUP98-HOXA9/N51S (NH9/N51S), HOXA9 (H9), or HOXA9ΔN (H9ΔN). Cells were sorted for GFP positivity and total RNA was subjected to microarray analysis. The experiment was performed two independent times and only genes that showed up- or down-regulation by 1.74 fold or more compared to control in both experiments were considered dysregulated. As shown in the upper Venn diagram, 46 probe sets showed dysregulation by both NUP98-HOXA9 (NH9) and NUP98-HOXA9/N51S (NH9/N51S). Of these, 26 did not show dysregulation by either HOXA9ΔN (H9ΔN) or HOXA9 (H9) and were considered homeodomain-independent (lower Venn diagram). These 26 probe sets represent 23 unique genes listed on the left. Genes with a potential role in homeodomain-independent transforming functions of NUP98-HOXA9 are highlighted in red and are discussed in the text. (B) Chromatin was immunoprecipitated with anti-Flag antibody from K562 cells nucleofected with either pTracer/CMV-Bsd vector (control), or vector containing Flag-tagged NUP98-HOXA9 (NH9) or NUP98-HOXA9/N51S (NH9/N51S). A segment of the HEY1 promoter was amplified by PCR (see Materials and Methods for primer sequences). The upper lanes show immunoprecipitated chromatin (IP) and the lower lanes show input chromatin (Input).