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. 2016 May 9;7(23):35341–35352. doi: 10.18632/oncotarget.9241

Figure 2. Inhibition of HOXA gene transcription in the presence of IRX1.

Figure 2

Stable HEK293T cell lines expressing IRX1 and/or MLL-AF4 were established to investigate HOXA gene transcription. A. Transcription of all transgenes after induction with doxycyline for two days (n=3). All transgenes were transcribed as expected. B. HOXA gene transcription levels (mock-transfected cells set to 1, normalized to GAPDH) in the presence of either IRX1 (light grey), MLL-AF4 (black) or IRX1/MLL-AF4 expressing cells (dark grey)(n=3). Except for HOXA7, all tested HOXA genes were downregulated in the presence of IRX1 and upregulated upon MLL-AF4 expression. Co-expression of IRX1 and MLL-AF4 leads to a downregulation of HOXA gene transcription. C. Validation of the effect of applied TSA administration. The control gene GAPDH used for Q-PCR experiments and the expression of the two induced transgenes were not affected. D. Same experiment as shown in B., however, with 300 nM TSA (n=3). E. Simple scheme to summarize that IRX1 acts dominant over MLL-AF4, while TSA blocks the action of IRX1 and MLL-AF4 while concomitantly activating the endogenous MLL.