Figure 4
From: Polycomb repressor complex 2 regulates HOXA9 and HOXA10, activating ID2 in NK/T-cell lines
Expression analysis in T-ALL cells. (A) JURKAT cells were lentivirally transduced for overexpression of HOXA9, HOXA10 and HOP, respectively. Subsequent RQ-PCR analysis of ID2 and RUNX2 demonstrated elevated ID2 transcription in HOXA9/HOXA10 overexpressing cells and reduced levels in HOP overexpressing cells. Expression levels of RUNX2 showed no alterations. (B) Western blot analysis demonstrated elevated expression of ID2 protein in JURKAT cells overexpressing HOXA10. STAT5 protein analysis served as loading control. (C) ChIP-PCR analysis in LOUCY (HOXA10-positive) and PEER (HOXA10-negative), using HOXA10 antibody demonstrated direct binding of HOXA10 to the ID2 promoter region at -2167 bp. (D) HELA cells were cotransfected with an ID2 promoter-construct and an expression-construct for HOXA10 or vector control. Reportergene activity was subsequently quantified by real-time PCR, indicating an activatory input by HOXA10.