Figure 5.

Effects of AML1/ETO and hypoxia on TRAIL promoter. (a) Portion of TRAIL promoter (180 bp, exon 1) containing the possible consensus sequence (box) for AML1. (b) U937-A/E cells were incubated for 2 days (from day −2 to time 0) in normoxia, in the absence (–) or the presence (+) of 5 μM ponasterone added daily (PON) and lysed. Lysates were subjected to ChIP using antibodies against ETO, RNApol II or control IgG, and then to RT-PCR for the TRAIL promoter (pTRAIL). (c) U937-A/E cells treated (PON) or not (NT) with ponasterone like in (b), or Kasumi-1 cells incubated for 3 days in normoxia (N) or hypoxia (H) were lysed and Q-PCR for TRAIL mRNA was then performed. Histograms report the relative quantification of TRAIL mRNA (normalized for GAPDH) with respect to ponasterone-treated U937-A/E cells (used as calibrator). (d) Kasumi-1 cells were incubated in normoxia (N) or hypoxia (H) for 3 days, lysed, lysates subjected to ChIP using the indicated antibodies and Q-PCR for TRAIL promoter (pTRAIL) then performed. (c, d) Histograms represent the average±S.E.M. of data from three independent experiments